Hepatitis C virus inhibitors

ABSTRACT

Hepatitis C virus inhibitors are disclosed having the general formula: 
     
       
         
         
             
             
         
       
         
         
           
             wherein R 1 , R 2 , R 3 , R′, B, Y and X are described in the description. Compositions comprising the compounds and methods for using the compounds to inhibit HCV are also disclosed.

CROSS REFERENCE TO RELATED APPLICATION

This continuation application claims the benefit of U.S. Ser. No.12/966,175 filed Dec. 13, 2010, now allowed, which in turn is acontinuation application which claims the benefit of U.S. Ser. No.12/202,603 filed Sep. 2, 2008, now U.S. Pat. No. 7,915,291, which inturn is a continuation application which claims the benefit of U.S. Ser.No. 11/295,914, filed Dec. 7, 2005, now U.S. Pat. No. 7,449,479, whichin turn is a continuation which claims the benefit of U.S. Ser. No.10/441,657, filed May 20, 2003, now U.S. Pat. No. 6,995,174, which inturn claims the benefit of provisional application U.S. Ser. No.60/382,055, filed May 20, 2002.

FIELD OF THE INVENTION

The present invention is generally directed to antiviral compounds, andmore specifically directed to compounds which inhibit the functioning ofthe NS3 protease encoded by Hepatitis C virus (HCV), compositionscomprising such compounds and methods for inhibiting the functioning ofthe NS3 protease.

BACKGROUND OF THE INVENTION

HCV is a major human pathogen, infecting an estimated 170 millionpersons worldwide—roughly five times the number infected by humanimmunodeficiency virus type 1. A substantial fraction of these HCVinfected individuals develop serious progressive liver disease,including cirrhosis and hepatocellular carcinoma. (Lauer, G. M.; Walker,B. D. N. Engl. J. Med. (2001), 345, 41-52).

Presently, the most effective HCV therapy employs a combination ofalpha-interferon and ribavirin, leading to sustained efficacy in 40% ofpatients. (Poynard, T. et al. Lancet (1998), 352, 1426-1432). Recentclinical results demonstrate that pegylated alpha-interferon is superiorto unmodified alpha-interferon as monotherapy (Zeuzem, S. et al. N.Engl. J. Med. (2000), 343, 1666-1672). However, even with experimentaltherapeutic regimens involving combinations of pegylatedalpha-interferon and ribavirin, a substantial fraction of patients donot have a sustained reduction in viral load. Thus, there is a clear andlong-felt need to develop effective therapeutics for treatment of HCVinfection.

HCV is a positive-stranded RNA virus. Based on a comparison of thededuced amino acid sequence and the extensive similarity in the 5′untranslated region, HCV has been classified as a separate genus in theFlaviviridae family. All members of the Flaviviridae family haveenveloped virions that contain a positive stranded RNA genome encodingall known virus-specific proteins via translation of a single,uninterrupted, open reading frame.

Considerable heterogeneity is found within the nucleotide and encodedamino acid sequence throughout the HCV genome. At least six majorgenotypes have been characterized, and more than 50 subtypes have beendescribed. The major genotypes of HCV differ in their distributionworldwide, and the clinical significance of the genetic heterogeneity ofHCV remains elusive despite numerous studies of the possible effect ofgenotypes on pathogenesis and therapy.

The single strand HCV RNA genome is approximately 9500 nucleotides inlength and has a single open reading frame (ORF) encoding a single largepolyprotein of about 3000 amino acids. In infected cells, thispolyprotein is cleaved at multiple sites by cellular and viral proteasesto produce the structural and non-structural (NS) proteins. In the caseof HCV, the generation of mature non-structural proteins (NS2, NS3,NS4A, NS4B, NS5A, and NS5B) is effected by two viral proteases. Thefirst one, as yet poorly characterized, cleaves at the NS2-NS3 junction;the second one is a serine protease contained within the N-terminalregion of NS3 (henceforth referred to as NS3 protease) and mediates allthe subsequent cleavages downstream of NS3, both in cis, at the NS3-NS4Acleavage site, and in trans, for the remaining NS4A-NS4B, NS4B-NS5A,NS5A-NS5B sites. The NS4A protein appears to serve multiple functions,acting as a cofactor for the NS3 protease and possibly assisting in themembrane localization of NS3 and other viral replicase components. Thecomplex formation of the NS3 protein with NS4A seems necessary to theprocessing events, enhancing the proteolytic efficiency at all of thesites. The NS3 protein also exhibits nucleoside triphosphatase and RNAhelicase activities. NS5B is a RNA-dependent RNA polymerase that isinvolved in the replication of HCV.

Among the compounds that have demonstrated efficacy in inhibiting HCVreplication, as selective HCV serine protease inhibitors, are thepeptide compounds disclosed in U.S. Pat. No. 6,323,180.

SUMMARY OF THE INVENTION

The present invention provides a compound of formula I, includingpharmaceutically acceptable salts, solvates or prodrugs thereof,

wherein:

-   -   (a) R₁ is C₁₋₈ alkyl, C₃₋₇ cycloalkyl, or C₄₋₁₀ alkylcycloalkyl;    -   (b) m is 1 or 2;    -   (c) n is 1 or 2;    -   (d) R₂ is H C₁₋₆ alkyl, C₂₋₆ alkenyl or C₃₋₇ cycloalkyl, each        optionally substituted with halogen;    -   (e) R₃ is C₁₋₈ alkyl optionally substituted with halo, cyano,        amino, C₁₋₆ dialkylamino, C₆₋₁₀ aryl, C₇₋₁₄ alkylaryl, C₁₋₆        alkoxy, carboxy, hydroxy, aryloxy, C₇₋₁₄ alkylaryloxy, C₂₋₆        alkylester, C₈₋₁₅ alkylarylester; C₃₋₁₂ alkenyl, C₃₋₇        cycloalkyl, or C₄₋₁₀ alkylcycloalkyl, wherein the cycloalkyl or        alkylcycloalkyl are optionally substituted with hydroxy, C₁₋₆        alkyl, C₂₋₆ alkenyl or C₁₋₆ alkoxy; or R₃ together with the        carbon atom to which it is attached forms a C₃₋₇ cycloalkyl        group optionally substituted with C₂₋₆ alkenyl;    -   (f) Y is H, phenyl substituted with nitro, pyridyl substituted        with nitro, or C₁₋₆ alkyl optionally substituted with cyano, OH        or C₃₋₇ cycloalkyl; provided that if R₄ or R₅ is H then Y is H;    -   (g) B is H, C₁₋₆ alkyl, R₄—(C═O)—, R₄O(C═O)—, R₄—N(R₅)—C(═O)—,        R₄—N(R₅)—C(═S)—, R₄SO₂—, or R₄—N(R₅)—SO₂—;    -   (h) R₄ is (i) C₁₋₁₀ alkyl optionally substituted with phenyl,        carboxyl, C₁₋₆ alkanoyl, 1-3 halogen, hydroxy, —OC(O)C₁₋₆ alkyl,        C₁₋₆ alkoxy, amino optionally substituted with C₁₋₆ alkyl,        amido, or (lower alkyl)amido; (ii) C₃₋₇cycloalkyl,        C₃₋₇cycloalkoxy, or C₄₋₁₀ alkylcycloalklyl, each optionally        substituted with hydroxy, carboxyl, (C₁₋₆ alkoxy)carbonyl, amino        optionally substituted with C₁₋₆ alkyl, amido, or (lower        alkyl)amido; (iii) C₆₋₁₀ aryl or C₇₋₁₆ arylalkyl, each        optionally substituted with C₁₋₆alkyl, halogen, nitro, hydroxy,        amido, (lower alkyl)amido, or amino optionally substituted with        C₁₋₆ alkyl; (iv) Het; (v) bicyclo(1.1.1)pentane; or (vi)        —C(O)OC₁₋₆alkyl, C₂₋₆alkenyl or C₂₋₆ alkynyl;    -   (i) R₅ is H; C₁₋₆ alkyl optionally substituted with 1-3        halogens; or C₁₋₆ alkoxy provided R₄ is C₁₋₁₀ alkyl;    -   (j) X is O, S, SO, SO₂, OCH₂, CH₂O or NH;    -   (k) R′ is Het; or C₆₋₁₀ aryl or C₇₋₁₄ alkylaryl, optionally        substituted with R^(a); and    -   (l) R^(a) is C₁₋₆ alkyl, C₃₋₇ cycloalkyl, C₁₋₆ alkoxy, C₃₋₇        cycloalkoxy, halo-C₁₋₆ alkyl, CF₃, mono- or di-halo-C₁₋₆ alkoxy,        cyano, halo, thioalkyl, hydroxy, alkanoyl, NO₂, SH, amino, C₁₋₆        alkylamino, di(C₁₋₆)alkylamino, di(C₁₋₆)alkylamide, carboxyl,        (C₁₋₆) carboxyester, C₁₋₆alkylsulfone, C₁₋₆ alkylsulfonamide,        di(C₁₋₆)alkyl(alkoxy)amine, C₆₋₁₀ aryl, C₇₋₁₄ alkylaryl, or a        5-7 membered monocyclic heterocycle;        with the proviso that X—R′ is not

or a pharmaceutically acceptable salt, solvate or prodrug thereof.

The present invention also provides compositions comprising thecompounds or pharmaceutically acceptable salts, solvates or prodrugsthereof and a pharmaceutically acceptable carrier. In particular, thepresent invention provides pharmaceutical compositions useful forinhibiting HCV NS3 comprising a therapeutically effective amount of acompound of the present invention, or a pharmaceutically acceptablesalt, solvate or prodrug thereof, and a pharmaceutically acceptablecarrier.

The present invention further provides methods for treating patientsinfected with HCV, comprising administering to the patient atherapeutically effective amount of a compound of the present invention,or a pharmaceutically acceptable salt, solvate or prodrug thereof.Additionally, the present invention provides methods of inhibiting HCVNS3 protease by administering to a patient an effective amount of acompound of the present invention.

By virtue of the present invention, it is now possible to provideimproved drugs comprising the compounds of the invention which can beeffective in the treatment of patients infected with HCV. Specifically,the present invention provides peptide compounds that can inhibit thefunctioning of the NS3 protease, e.g., in combination with the NS4Aprotease.

DETAILED DESCRIPTION OF THE INVENTION

Stereochemical definitions and conventions used herein generally followMcGraw-Hill Dictionary of Chemical Terms, S. P. Parker, Ed., McGraw-HillBook Company, New York (1984) and Stereochemistry of Organic Compounds,Eliel, E. and Wilen, S., John Wiley & Sons, Inc., New York (1994). Manyorganic compounds exist in optically active forms, i.e., they have theability to rotate the plane of plane-polarized light. In describing anoptically active compound, the prefixes D and L or R and S are used todenote the absolute configuration of the molecule about its chiralcenter(s). The prefixes d and l or (+) and (−) are employed to designatethe sign of rotation of plane-polarized light by the compound, with (−)or 1 meaning that the compound is levorotatory and (+) or d, meaning thecompound, is dextrorotatory. For a given chemical structure, thesecompounds, called stereoisomers, are identical except that they aremirror images of one another. A specific stereoisomer of a mirror imagepair may also be referred to as an enantiomer, and a mixture of suchisomers is often called an enantiomeric mixture.

The nomenclature used to describe organic radicals, e.g., hydrocarbonsand substituted hydrocarbons, generally follows standard nomenclatureknown in the art, unless otherwise specifically defined. Combinations ofgroups, e.g., alkylalkoxyamine, include all possible stableconfigurations, unless otherwise specifically stated. Certain radicalsand combinations are defined below for purposes of illustration.

The terms “racemic mixture” and “racemate” refer to an equimolar mixtureof two enantiomeric species, devoid of optical activity.

The term “chiral” refers to molecules which have the property ofnon-superimposability of the mirror image partner, while the term“achiral” refers to molecules which are superimposable on their mirrorimage partner.

The term “stereoisomers” refers to compounds which have identicalchemical composition, but differ with regard to the arrangement of theatoms or groups in space.

The term “diastereomer” refers to a stereoisomer which is not anenantiomer, e.g., a stereoisomer with two or more centers of chiralityand whose molecules are not mirror images of one another. Diastereomershave different physical properties, e.g. melting points, boiling points,spectral properties, and reactivities. Mixtures of diastereomers mayseparate under high resolution analytical procedures such aselectrophoresis and chromatography.

The term “enantiomers” refers to two stereoisomers of a compound whichare non-superimposable mirror images of one another.

The term “pharmaceutically acceptable salt” is intended to includenontoxic salts synthesized from a compound which contains a basic oracidic moiety by conventional chemical methods. Generally, such saltscan be prepared by reacting the free acid or base forms of thesecompounds with a stoichiometric amount of the appropriate base or acidin water or in an organic solvent, or in a mixture of the two;generally, nonaqueous media like ether, ethyl acetate, ethanol,isopropanol, or acetonitrile are preferred. Lists of suitable salts arefound in Remington's Pharmaceutical Sciences, 18th ed., Mack PublishingCompany, Easton, Pa., 1990, p. 1445. The compounds of the presentinvention are useful in the form of the free base or acid or in the formof a pharmaceutically acceptable salt thereof. All forms are within thescope of the invention.

The term “therapeutically effective amount” means the total amount ofeach active component that is sufficient to show a meaningful patientbenefit, e.g., a sustained reduction in viral load. When applied to anindividual active ingredient, administered alone, the term refers tothat ingredient alone. When applied to a combination, the term refers tocombined amounts of the active ingredients that result in thetherapeutic effect, whether administered in combination, serially orsimultaneously.

The term “compounds of the invention”, and equivalent expressions, aremeant to embrace compounds of Formula I, and pharmaceutically acceptablesalts, and solvates, e.g. hydrates. Similarly, reference tointermediates, is meant to embrace their salts, and solvates, where thecontext so permits. References to the compound of the invention alsoinclude the preferred compounds of Formula II and III.

The term “derivative” means a chemically modified compound wherein themodification is considered routine by the ordinary skilled chemist, suchas an ester or an amide of an acid, protecting groups, such as a benzylgroup for an alcohol or thiol, and tert-butoxycarbonyl group for anamine.

The term “solvate” means a physical association of a compound of thisinvention with one or more solvent molecules, whether organic orinorganic. This physical association includes hydrogen bonding. Incertain instances the solvate will be capable of isolation, for examplewhen one or more solvent molecules are incorporated in the crystallattice of the crystalline solid. “Solvate” encompasses bothsolution-phase and isolable solvates. Exemplary solvates includehydrates, ethanolates, methanolates, and the like.

The term “prodrug” as used herein means derivatives of the compounds ofthe invention which have chemically or metabolically cleavable groupsand become, by solvolysis or under physiological conditions, thecompounds of the invention which are pharmaceutically active in vivo. Aprodrug of a compound may be formed in a conventional manner with afunctional group of the compounds such as with an amino, hydroxy orcarboxy group when present. The prodrug derivative form often offersadvantages of solubility, tissue compatibility, or delayed release in amammalian organism (see, Bundgard, H., Design of Prodrugs, pp. 7-9,21-24, Elsevier, Amsterdam 1985). Prodrugs include acid derivatives wellknown to practitioners of the art, such as, for example, esters preparedby reaction of the parent acidic compound with a suitable alcohol, oramides prepared by reaction of the parent acid compound with a suitableamine.

The term “patient” includes both human and other mammals.

The term “pharmaceutical composition” means a composition comprising acompound of the invention in combination with at least one additionalpharmaceutical carrier, i.e., adjuvant, excipient or vehicle, such asdiluents, preserving agents, fillers, flow regulating agents,disintegrating agents, wetting agents, emulsifying agents, suspendingagents, sweetening agents, flavoring agents, perfuming agents,antibacterial agents, antifungal agents, lubricating agents anddispensing agents, depending on the nature of the mode of administrationand dosage forms. Ingredients listed in Remington's PharmaceuticalSciences, 18^(th) ed., Mack Publishing Company, Easton, Pa. (1999) forexample, may be used.

The phrase “pharmaceutically acceptable” is employed herein to refer tothose compounds, materials, compositions, and/or dosage forms which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of patients without excessive toxicity, irritation,allergic response, or other problem or complication commensurate with areasonable risk/benefit ratio.

The term “treating” refers to: (i) preventing a disease, disorder orcondition from occurring in a patient which may be predisposed to thedisease, disorder and/or condition but has not yet been diagnosed ashaving it; (ii) inhibiting the disease, disorder or condition, i.e.,arresting its development; and (iii) relieving the disease, disorder orcondition, i.e., causing regression of the disease, disorder and/orcondition. The term “substituted” as used herein includes substitutionat from one to the maximum number of possible binding sites on the core,e.p., organic radical, to which the substitutent is bonded, e.g., mono-,di-, tri- or tetra-substituted, unless otherwise specifically stated.

The term “halo” as used herein means a halogen substituent selected frombromo, chloro, fluoro or iodo. The term “haloalkyl” means an alkyl groupthat in substituted with one or more halo substituents.

The term “alkyl” as used herein means acyclic, straight or branchedchain alkyl substituents and includes, for example, methyl, ethyl,propyl, butyl, tert-butyl, hexyl, 1-methylethyl, 1-methylpropyl,2-methylpropyl, 1,1-dimethylethyl. Thus, C₁₋₆ alkyl refers to an alkylgroup having from one to six carbon atoms. The term “lower alkyl” meansan alkyl group having from one to six, preferably from one to fourcarbon atoms. The term “alkylester” means an alkyl group additionallycontaining on ester group. Generally, a stated carbon number range,e.g., C₂₋₆ alkylester, includes all of the carbon atoms in the radical.

The term “alkenyl” as used herein means an alkyl radical containing atleast one double bond, e.g., ethenyl(vinyl) and alkyl.

The term “alkoxy” as used herein means an alkyl group with the indicatednumber of carbon atoms attached to an oxygen atom. Alkoxy includes, forexample, methoxy, ethoxy, propoxy, 1-methylethoxy, butoxy and1,1-dimethylethoxy. The latter radical is referred to in the art astert-butoxy. The term “alkoxycarbonyl” means an alkoxy groupadditionally containing a carbonyl group.

The term “cycloalkyl” as used herein means a cycloalkyl substituentcontaining the indicated number of carbon atoms and includes, forexample, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyland spiro cyclic groups such as spirocyclopropyl as spirocyclobutyl. Theterm “cycloalkoxy” as used herein means a cycloalkyl group linked to anoxygen atom, such as, for example, cyclobutyloxy or cyclopropyloxy. Theterm “alkylcycloalkyl” means a cycloalkyl group linked to an alkylgroup. The stated carbon number range includes the total number ofcarbons in the radical, unless otherwise specifically stated. This aC₄₋₁₀ alkylcycloalkyl may contain from 1-7 carbon atoms in the alkylgroup and from 3-9 carbon atoms in the ring, e.g., cyclopropylmethyl orcyclohexylethyl.

The term “aryl” as used herein means an aromatic moiety containing theindicated number of carbon atoms, such as, but not limited to phenyl,indanyl or naphthyl. For example, C₆₋₁₀ aryl refers to an aromaticmoiety having from six to ten carbon atoms which may be in the form of amonocyclic or bicyclic structure. The term “haloaryl” as used hereinrefers to an aryl mono, di or tri substituted with one or more halogenatoms. The terms “alkylaryl”, “arylalkyl” and “aralalkyl” mean an arylgroup substituted with one or more alkyl groups. Thus, a C₇₋₁₄ alkylarylgroup many have from 1-8 carbon atoms in the alkyl group for amonocyclic aromatic and from 1-4 carbon atoms in the alkyl group for afused aromatic. The aryl radicals include those substituted with typicalsubstituents known to those skilled in the art, e.g., halogen, hydroxy,carboxy, carbonyl, nitro, sulfo, amino, cyano, dialkylamino haloalkyl,CF₃, haloalkoxy, thioalkyl, alkanoyl, SH, alkylamino, alkylamide,dialkylamide, carboxyester, alkylsulfone, alkylsulfonamide andalkyl(alkoxy)amine. Examples of alkylaryl groups include benzyl,butylphenyl and 1-naphthylmethyl. The terms “alkylaryloxy” and“alkylarylester” mean alkylaryl groups containing an oxygen atom andester group, respectively.

The term “carboxyalkyl” as used herein means a carboxyl group (COOH)linked through an alkyl group as defined above and includes, forexample, butyric acid.

The term “alkanoyl” as used herein means straight or branched 1-oxoalkylradicals containing the indicated number of carbon atoms and includes,for example, formyl, acetyl, 1-oxopropyl(propionyl),2-methyl-1-oxopropyl, 1-oxohexyl and the like.

The term “amino aralkyl” as used herein means an amino group substitutedwith an aralkyl group, such as the following amino aralkyl

The term “alkylamide” as used herein means an amide mono-substitutedwith an alkyl, such as

The term “carboxyalkyl” as used herein means a carboxyl group (COOH)linked through a alkyl group as defined above and includes, for example,butyric acid.

The term “heterocycle”, also referred to as “Het”, as used herein means7-12 membered bicyclic heterocycles and 5-7 membered monocyclicheterocycles.

Preferred bicyclic heterocycles are 7-12 membered fused bicyclic ringsystems (both rings share an adjacent pair of atoms) containing from oneto four heteroatoms selected from nitrogen, oxygen and sulfur, whereinboth rings of the heterocycle are fully unsaturated. The nitrogen andsulfur heteroatoms atoms may be optionally oxidized. The bicyclicheterocycle may contain the heteroatoms in one or both rings. Thebicyclic heterocycle may also contain substituents on any of the ringcarbon atoms, e.g., one to three substituents. Examples of suitablesubstituents include C₁₋₆ alkyl, C₃₋₇ cycloalkyl, C₁₋₆ alkoxy, C₃₋₇cycloalkoxy, halo-C₁₋₆ alkyl, CF₃, mono- or di-halo-C₁₋₆ alkoxy, cyano,halo, thioalkyl, hydroxy, alkanoyl, NO₂, SH, amino, C₁₋₆ alkylamino,di(C₁₋₆)alkylamino, di(C₁₋₆)alkylamide, carboxyl, (C₁₋₆) carboxyester,C₁₋₆ alkylsulfone, C₁₋₆ alkylsulfonamide, C₁₋₆ alkylsulfoxide,di(C₁₋₆)alkyl(alkoxy)amine, C₆₋₁₀ aryl, C₇₋₁₄ alkylaryl, and a 5-7membered monocyclic heterocycle. When two substituents are attached tovicinal carbon atoms of the bicyclic heterocycle, they can join to forma ring, e.g., a five, six or seven membered ring system containing up totwo heteroatoms selecting from oxygen and nitrogen. The bicyclicheterocycle may be attached to its pendant group, e.g. X in Formula I,at any atom in the ring and preferably carbon.

Examples of bicyclic heterocycles include, but are not limited to, thefollowing ring systems:

Preferred monocyclic heterocycles are 5-7 membered saturated, partiallysaturated or fully unsaturated ring system (this latter subset hereinreferred to as unsaturated heteroaromatic) containing in the ring fromone to four heteroatoms selected from nitrogen, oxygen and sulfur,wherein the sulfur and nitrogen heteroatoms may be optionally oxidized.The monocyclic heterocycle may also contain substituents on any of thering atoms, e.g., one to three substituents. Examples of suitablesubstituents include C₁₋₆ alkyl, C₃₋₇ cycloalkyl, C₁₋₆ alkoxy, C₃₋₇cycloalkoxy, halo-C₁₋₆ alkyl, CF₃, mono- or di-halo-C₁₋₆ alkoxy, cyano,halo, thioalkyl, hydroxy, alkanoyl, NO₂, SH, amino, C₁₋₆ alkylamino,di(C₁₋₆)alkylamino, di(C₁₋₆)alkylamide, carboxyl, (C₁₋₆) carboxyester,C₁₋₆ alkylsulfone, C₁₋₆ alkylsulfoxide, C₁₋₆ alkylsulfonamide,di(C₁₋₆)alkyl(alkoxy)amine, C₆₋₁₀ aryl, C₇₋₁₄ alkylaryl and 5-7 memberedmonocyclic heterocycle. The monocyclic heterocycle may be attached toits pendant group, e.g. X in Formula I, at any atom in the ring.

Examples of monocyclic heterocycles include, but are not limited to, thefollowing:

Those skilled in the art will recognize that the heterocycles used inthe compounds of the present invention should be stable. Generally,stable compounds are those which can be synthesized, isolated andformulated using techniques known the those skilled in the art withoutdegradation of the compound.

Where used in naming compounds of the present invention, thedesignations “P1′, P1, P2, P3 and P4”, as used herein, map the relativepositions of the amino acid residues of a protease inhibitor bindingrelative to the binding of the natural peptide cleavage substrate.Cleavage occurs in the natural substrate between P1 and P1′ where thenonprime positions designate amino acids starting from the C-terminusend of the peptide natural cleavage site extending towards theN-terminus; whereas, the prime positions emanate from the N-terminus endof the cleavage site designation and extend towards the C-terminus. Forexample, P1′ refers to the first position away from the right hand endof the C-terminus of the cleavage site (ie. N-terminus first position);whereas P1 starts the numbering from the left hand side of theC-terminus cleavage site, P2: second position from the C-terminus,etc.)(see Berger A. & Schechter I., Transactions of the Royal SocietyLondon series (1970), B257, 249-264].

Thus in the compounds of formula I, the “P1′ to P4” portions of themolecule are indicated below:

As used herein the term “1-aminocyclopropyl-carboxylic acid” (Acca)refers to a compound of formula:

As used herein the term “tert-butylglycine” refers to a compound of theformula:

The term “residue” with reference to an amino acid or amino acidderivative means a radical derived from the corresponding α-amino acidby eliminating the hydroxyl of the carboxy group and one hydrogen of theα-amino acid group. For instance, the terms Gln, Ala, Gly, Ile, Arg,Asp, Phe, Ser, Leu, Cys, Asn, Sar and Tyr represent the “residues” ofL-glutamine, L-alanine, glycine, L-isoleucine, L-arginine, L-asparticacid, L-phenylalanine, L-serine, L-leucine, L-cysteine, L-asparagine,sarcosine and L-tyrosine, respectively.

The term “side chain” with reference to an amino acid or amino acidresidue means a group attached to the α-carbon atom of the α-amino acid.For example, the R-group side chain for glycine is hydrogen, for alanineit is methyl, for valine it is isopropyl. For the specific R-groups orside chains of the α-amino acids reference is made to A. L. Lehninger'stext on Biochemistry (see chapter 4).

The compounds of the present invention have the structure of Formula I:

wherein:

-   -   (a) R₁ is C₁₋₈alkyl, C₃₋₇ cycloalkyl, or C₄₋₁₀ alkylcycloalkyl;    -   (b) m is 1 or 2;    -   (c) n is 1 or 2;    -   (d) R₂ is H C₁₋₆ alkyl, C₂₋₆ alkenyl or C₃₋₇ cycloalkyl, each        optionally substituted with halogen;    -   (e) R₃ is C₁₋₈ alkyl optionally substituted with halo, cyano,        amino, C₁₋₆ dialkylamino, C₆₋₁₀ aryl, C₇₋₁₄ alkylaryl, C₁₋₆        alkoxy, carboxy, hydroxy, aryloxy, C₇₋₁₄ alkylaryloxy, C₂₋₆        alkylester, C₈-15 alkylarylester; C₃₋₁₂ alkenyl, C₃₋₇        cycloalkyl, or C₄₋₁₀ alkylcycloalkyl, wherein the cycloalkyl or        alkylcycloalkyl are optionally substituted with hydroxy, C₁₋₆        alkyl, C₂₋₆ alkenyl or C₁₋₆ alkoxy; or R₃ together with the        carbon atom to which it is attached forms a C₃₋₇ cycloalkyl        group optionally substituted with C₂₋₆ alkenyl;    -   (f) Y is H, phenyl substituted with nitro, pyridyl substituted        with nitro, or C₁₋₆ alkyl optionally substituted with cyano, OH        or C₃₋₇ cycloalkyl; provided that if R₄ or R₅ is H then Y is H;    -   (g) B is H, C₁₋₆ alkyl, R₄—(C═O)—, R₄O(C═O)—, R₄—N(R₅)—C(═O)—,        R₄—N(R₅)—C(═S)—, R₄SO₂—, or R₄—N(R₅)—SO₂—;    -   (h) R₄ is (i) C₁₋₁₀ alkyl optionally substituted with phenyl,        carboxyl, C₁₋₆ alkanoyl, 1-3 halogen, hydroxy, —OC(O)C₁₋₆ alkyl,        C₁₋₆ alkoxy, amino optionally substituted with C₁₋₆ alkyl,        amido, or (lower alkyl)amido; (ii) C₃₋₇ cycloalkyl, C₃₋₇        cycloalkoxy, or C₄₋₁₀ alkylcycloalklyl, each optionally        substituted with hydroxy, carboxyl, (C₁₋₆ alkoxy)carbonyl, amino        optionally substituted with C₁₋₆ alkyl, amido, or (lower        alkyl)amido; (iii) C₆₋₁₀ aryl or C₇₋₁₆ arylalkyl, each        optionally substituted with C₁₋₆alkyl, halogen, nitro, hydroxy,        amido, (lower alkyl)amido, or amino optionally substituted with        C₁₋₆ alkyl; (iv) Het; (v) bicyclo(1.1.1)pentane; or (vi)        —C(O)OC₁₋₆ alkyl, C₂₋₆ alkenyl or C₂₋₆alkynyl;    -   (i) R₅ is H; C₁₋₆ alkyl optionally substituted with 1-3        halogens; or C₁₋₆ alkoxy provided R₄ is C₁₋₁₀ alkyl;    -   (j) X is O, S, SO, SO₂, OCH₂, CH₂O or NH;    -   (k) R′ is Het; or C₆₋₁₀ aryl or C₇₋₁₄ alkylaryl, optionally        substituted with R^(a); and    -   (l) R^(a) is C₁₋₆ alkyl, C₃₋₇ cycloalkyl, C₁₋₆ alkoxy, C₃₋₇        cycloalkoxy, halo-C₁₋₆ alkyl, CF₃, mono- or di-halo-C₁₋₆ alkoxy,        cyano, halo, thioalkyl, hydroxy, alkanoyl, NO₂, SH, amino, C₁₋₆        alkylamino, di(C₁₋₆)alkylamino, di(C₁₋₆)alkylamide, carboxyl,        (C₁₋₆) carboxyester, C₁₋₆ alkylsulfone, C₁₋₆ alkylsulfonamide,        di(C₁₋₆)alkyl(alkoxy)amine, C₆₋₁₀ aryl, C₇₋₁₄ alkylaryl, or a        5-7 membered monocyclic heterocycle;        with the proviso that X—R′ is not

or a pharmaceutically acceptable salt, solvate or prodrug thereof.

Preferably, R₂ is C₂₋₆ alkenyl; R₃ is C₁₋₈ alkyl optionally substitutedwith C₁₋₆ alkoxy, or C₃₋₇ cycloalkyl; Y is H; B is R₄—(C═O)—, R₄O(C═O)—,or R₄—N(R₅)—C(═O)—; R₄ is C₁₋₁₀ alkyl optionally substituted with 1-3halogen or C₁₋₆ alkoxy; or C₃₋₇ cycloalkyl or C₄₋₁₀ alkylcycloalklyl; R₅is H; X is O or NH; and R′ is Het.

The substituents from each grouping may be selected individually andcombined in any combination which provides a stable compound inaccordance with the present invention. Also, more than one substituentfrom each group may be substituted on the core group provided there aresufficient available binding sites. For example, each of the followingR_(a) substituents, C₁₋₆ alkoxy, C₆ aryl and a 5-7 membered monocyclicheterocycle, may be substituted on a bicyclic heterocycle R′.

In a preferred aspect, the compounds of the present invention have thestructure of Formula II:

wherein:

-   -   (a) R₁ is C₃₋₇ cycloalkyl;    -   (b) R₂ is C₁₋₆ alkyl, C₂₋₆ alkenyl or C₃₋₇ cycloalkyl;    -   (c) R₃ is C₁₋₈ alkyl optionally substituted with C₆ aryl, C₁₋₆        alkoxy, carboxy, hydroxy, aryloxy, C₇₋₁₄ alkylaryloxy, C₂₋₆        alkylester, C₈₋₁₅ alkylarylester; C₃₋₁₂ alkenyl, C₃₋₇        cycloalkyl, or C₄₋₁₀ alkylcycloalkyl;    -   (d) Y is H;    -   (e) B is H, C₁₋₆ alkyl, R₄—(C═O)—, R₄O(C═O)—, R₄—N(R₅)—C(═O)—,        R₄—N(R₅)—C(═S)—, R₄SO₂—, or R₄—N(R₅)—SO₂—;    -   (f) R₄ is (i) C₁₋₁₀ alkyl optionally substituted with phenyl,        carboxyl, C₁₋₆ alkanoyl, 1-3 halogen, hydroxy, C₁₋₆ alkoxy; (ii)        C₃₋₇ cycloalkyl, C₃₋₇ cycloalkoxy, or C₄₋₁₀ alkylcycloalklyl;        or (iii) C₆₋₁₀ aryl or C₇₋₁₆ arylalkyl, each optionally        substituted with C₁₋₆ alkyl or halogen;    -   (g) R₅ is H or C₁₋₆ alkyl optionally substituted with 1-3        halogens;    -   (h) X is O or NH;    -   (i) R′ is Het; or C₆₋₁₀ aryl optionally substituted with R^(a);        and    -   (j) R^(a) is C₁₋₆ alkyl, C₃₋₇ cycloalkyl, C₁₋₆ alkoxy, halo-C₁₋₆        alkyl, halo, amino, C₆ aryl, or a 5-7 membered monocyclic        heterocycle;        with the proviso that X_(a)—R′ is not

or a pharmaceutically acceptable salt, solvate or prodrug thereof.

In one preferred aspect of the invention, R′ is a bicyclic heterocycle.Preferably, the bicyclic heterocycle contains 1 or 2 nitrogen atoms andoptionally a sulfur atom or an oxygen atom in the ring. Preferably, theheterocycle is substituted with at least one of C₁₋₆ alkyl, C₁₋₆ alkoxy,halo, C₆ aryl, and a 5-7 membered monocyclic heterocycle. Morepreferably, R′ is a bicyclic heterocycle containing 1 nitrogen atom andsubstituted with methoxy and at least one of a C₆ aryl and a 5-7membered monocyclic heterocycle.

In another preferred aspect of the invention, R′ is a monocyclicheterocycle. Preferably, the heterocycle contains 1 or 2 nitrogen atomsand optionally a sulfur atom or an oxygen atom in the ring. Preferably,the heterocycle is substituted with at least one of C₁₋₆ alkyl, C₁₋₆alkoxy, halo, C₆₋₁₀ aryl, C₇₋₁₄ alkylaryl, or a 5-7 membered monocyclicheterocycle. More preferably, R′ is a monoyclic heterocycle containing 1or 2 nitrogen atoms and substituted with methoxy and at least one of aC₆ aryl and a 5-7 membered monocyclic heterocycle.

In a more preferred aspect of the invention, the compounds have thestructure of Formula III

wherein:

-   -   (a) R₁ is C₃₋₇ cycloalkyl;    -   (b) R₂ is C₂₋₆ alkenyl;    -   (c) R₃ is C₁₋₈ alkyl;    -   (d) Y is H;    -   (e) B is R₄O(C═O)—, or R₄—N(R₅)—C(═O)—;    -   (f) R₄ is C₁₋₁₀ alkyl;    -   (g) R₅ is H;    -   (h) R′ is a bicyclic heterocycle optionally substituted with        R^(a); and    -   (i) R^(a) is C₁₋₆ alkyl, C₁₋₆ alkoxy, halo, C₆ aryl, or a 5-7        membered monocyclic heterocycle; with the proviso that O—R′ is        not OCH₃

-   -   -   or a pharmaceutically acceptable salt, solvate or prodrug            thereof.

Preferably, R₁ is cyclopropyl or cyclobutyl, R₂ is vinyl, R₃ is t-butyl,R₄ is t-butyl and R′ is quinoline or isoquinoline optionally substitutedwith R^(a). Preferably, R^(a) includes at least one of C₁₋₆ alkoxy, C₆aryl and a 5-7 membered monocyclic heterocycle. In a preferred aspect ofthe invention, R₁ is cyclopropyl, R₂ is vinyl, R₃ is t-butyl, R₄ ist-butyl, and R′ is isoquinoline substituted with C₁₋₆ alkoxy and atleast one of C₆ aryl or a 5-7 membered monocyclic heterocycle.

The compounds of the present invention, by virtue of their basic moiety,can form salts by the addition of a pharmaceutically acceptable acid.The acid addition salts are formed from a compound of Formula I and apharmaceutically acceptable inorganic acid, including but not limited tohydrochloric, hydrobromic, hydroiodic, sulfuric, phosphoric, or organicacid such as p-toluenesulfonic, methanesulfonic, acetic, benzoic,citric, malonic, fumaric, maleic, oxalic, succinic, sulfamic, ortartaric. Thus, examples of such pharmaceutically acceptable saltsinclude chloride, bromide, iodide, sulfate, phosphate, methanesulfonate,citrate, acetate, malonate, fumarate, sulfamate, and tartrate.

Salts of an amine group may also comprise quaternary ammonium salts inwhich the amino nitrogen carries a suitable organic group such as analkyl, alkenyl, alkynyl or aralkyl moiety.

Compounds of the present invention, which are substituted with an acidicgroup, may exist as salts formed through base addition. Such baseaddition salts include those derived from inorganic bases which include,for example, alkali metal salts (e.g. sodium and potassium), alkalineearth metal salts (e.g. calcium and magnesium), aluminum salts andammonium salts. In addition, suitable base addition salts include saltsof physiologically acceptable organic bases such as trimethylamine,triethylamine, morpholine, pyridine, piperidine, picoline,dicyclohexylamine, N,N′-dibenzylethylenediamine, 2-hydroxyethylamine,bis-(2-hydroxyethyl)amine, tri-(2-hydroxyethyl)amine, procaine,dibenzylpiperidine, N-benzyl-β-phenethylamine, dehydroabietylamine,N,N′-bishydroabietylamine, glucamine, N-methylglucamine, collidine,quinine, quinoline, ethylenediamine, ornithine, choline,N,N′-benzylphenethylamine, chloroprocaine, diethanolamine, diethylamine,piperazine, tris(hydroxymethyl)aminomethane and tetramethylammoniumhydroxide and basic amino acids such as lysine, arginine andN-methylglutamine. These salts may be prepared by methods known to thoseskilled in the art.

Certain compounds of the present invention, and their salts, may alsoexist in the form of solvates with water, for example hydrates, or withorganic solvents such as methanol, ethanol or acetonitrile to form,respectively, a methanolate, ethanolate or acetonitrilate. The presentinvention includes each solvate and mixtures thereof.

In addition, compounds of the present invention, or a salt or solvatethereof, may exhibit polymorphism. The present invention alsoencompasses any such polymorphic form.

The compounds of the present invention also contain two or more chiralcenters. For example, the compounds may include P1 cyclopropyl elementof formula

wherein C₁ and C₂ each represent an asymmetric carbon atom at positions1 and 2 of the cyclopropyl ring. Not withstanding other possibleasymmetric centers at other segments of the compounds, the presence ofthese two asymmetric centers means that the compounds can exist asracemic mixtures of diastereomers, such as the diastereomers wherein R₂is configured either syn to the amide or syn to the carbonyl as shownbelow.

The present invention includes both enantiomers and mixtures ofenantiomers such as racemic mixtures.

The enantiomers may be resolved by methods known to those skilled in theart, for example, by formation of diastereoisomeric salts which may beseparated by crystallization, gas-liquid or liquid chromatography,selective reaction of one enantiomer with an enantiomer-specificreagent. It will be appreciated that where the desired enantiomer isconverted into another chemical entity by a separation technique, thenan additional step is required to form the desired enantiomeric form.Alternatively, specific enantiomers may be synthesized by asymmetricsynthesis using optically active reagents, substrates, catalysts orsolvents, or by converting one enantiomer into the other by asymmetrictransformation.

The compounds of the present invention may be in the form of a prodrug.Simple aliphatic or aromatic esters derived from, when present, acidicgroups pendent on the compounds of this invention are preferredprodrugs. In some cases it is desirable to prepare double ester typeprodrugs such as (acyloxy)alkyl esters or (alkoxycarbonyl)oxy)alkylesters.

Certain compounds of the present invention may also exist in differentstable conformational forms which may be separable. Torsional asymmetrydue to restricted rotation about an asymmetric single bond, for examplebecause of steric hindrance or ring strain, may permit separation ofdifferent conformers. The present invention includes each conformationalisomer of these compounds and mixtures thereof.

Certain compounds of the present invention may exist in zwitterionicform and the present invention includes each zwitterionic form of thesecompounds and mixtures thereof.

The starting materials useful to synthesize the compounds of the presentinvention are known to those skilled in the art and can be readilymanufactured or are commercially available.

The compounds of the present invention can be manufactured by methodsknown to those skilled in the art, see e.p., U.S. Pat. No. 6,323,180 andUS Patent Appl. 20020111313 A1. The following methods set forth beloware provided for illustrative purposes and are not intended to limit thescope of the claimed invention. It will be recognized that it may bepreferred or necessary to prepare such a compound in which a functionalgroup is protected using a conventional protecting group then to removethe protecting group to provide a compound of the present invention. Thedetails concerning the use of protecting groups in accordance with thepresent invention are known to those skilled in the art.

The compounds of the present invention may, for example, be synthesizedaccording to a general process as illustrated in Scheme I (wherein CPGis a carboxyl protecting group and APG is an amino protecting group):

Briefly, the P1, P2, and P3 can be linked by well known peptide couplingtechniques. The P1, P2, and P3 groups may be linked together in anyorder as long as the final compound corresponds to peptides of theinvention. For example, P3 can be linked to P2-P1; or P1 linked toP3-P2.

Generally, peptides are elongated by deprotecting the α-amino group ofthe N-terminal residue and coupling the unprotected carboxyl group ofthe next suitably N-protected amino acid through a peptide linkage usingthe methods described. This deprotection and coupling procedure isrepeated until the desired sequence is obtained. This coupling can beperformed with the constituent amino acids in stepwise fashion, asdepicted in Scheme I.

Coupling between two amino acids, an amino acid and a peptide, or twopeptide fragments can be carried out using standard coupling proceduressuch as the azide method, mixed carbonic-carboxylic acid anhydride(isobutyl chloroformate) method, carbodiimide (dicyclohexylcarbodiimide,diisopropylcarbodiimide, or water-soluble carbodiimide) method, activeester (ρ-nitrophenyl ester, N-hydroxysuccinic imido ester) method,Woodward reagent K-method, carbonyldiimidazole method, phosphorusreagents or oxidation-reduction methods. Some of these methods(especially the carbodiimide method) can be enhanced by adding1-hydroxybenzotriazole or 4-DMAP. These coupling reactions can beperformed in either solution (liquid phase) or solid phase.

More explicitly, the coupling step involves the dehydrative coupling ofa free carboxyl of one reactant with the free amino group of the otherreactant in the present of a coupling agent to form a linking amidebond. Descriptions of such coupling agents are found in generaltextbooks on peptide chemistry, for example, M. Bodanszky, “PeptideChemistry”, 2^(nd) rev ed., Springer-Verlag, Berlin, Germany, (1993).Examples of suitable coupling agents are N,N′-dicyclohexylcarbodiimide,1-hydroxybenzotriazole in the presence of N,N′-dicyclohexylcarbodiimideor N-ethyl-N′-[(3-dimethylamino)propyl]carbodiimide. A practical anduseful coupling agent is the commercially available(benzotriazol-1-yloxy)tris-(dimethylamino)phosphoniumhexafluorophosphate, either by itself or in the present of1-hydroxybenzotriazole or 4-DMAP. Another practical and useful couplingagent is commercially available2-(1H-benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate.Still another practical and useful coupling agent is commerciallyavailable O-(7-azabenzotrizol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate. The coupling reaction is conducted in an inertsolvent, e.g. dichloromethane, acetonitrile or dimethylformamide. Anexcess of a tertiary amine, e.g. diisopropylethylamine,N-methylmorpholine, N-methylpyrrolidine or 4-DMAP is added to maintainthe reaction mixture at a pH of about 8. The reaction temperatureusually ranges between 0° C. and 50° C. and the reaction time usuallyranges between min and 24 h.

The functional groups of the constituent amino acids generally must beprotected during the coupling reactions to avoid formation of undesiredbonds. Protecting groups that can be used are listed, for example, inGreene, “Protective Groups in Organic Chemistry”, John Wiley & Sons, NewYork (1981) and “The Peptides: Analysis, Synthesis, Biology”, Vol. 3,Academic Press, New York (1981), the disclosures of which are herebyincorporated by reference.

The α-amino group of each amino acid to be coupled to the growingpeptide chain must be protected (APG). Any protecting group known in theart can be used. Examples of such groups include: 1) acyl groups such asformyl, trifluoroacetyl, phthalyl, and ρ-toluenesulfonyl; 2) aromaticcarbamate groups such as benzyloxycarbonyl (Cbz or Z) and substitutedbensyloxycarbonyls, and 9-fluorenylmethyloxycarbonyl (Fmoc); 3)aliphatic carbamate groups such as tert-butyloxycarbonyl (Boc),ethoxycarbonyl, diisopropylmethoxycarbonyl, and allyloxycarbonyl; 4)cyclic alkyl carbamate groups such as cyclopentyloxycarbonyl andadamantyloxycarbonyl; 5) alkyl groups such as triphenylmethyl andbenzyl; 6)trialkylsilyl such as trimethylsilyl; and 7) thiol containinggroups such as phenylthiocarbonyl and dithiasuccinoyl.

The preferred α-amino protecting group is either Boc or Fmoc. Many aminoacid derivatives suitably protected for peptide synthesis arecommercially available.

The α-amino protecting group of the newly added amino acid residue iscleaved prior to the coupling of the next amino acid. When the Boc groupis used, the methods of choice are trifluoroacetic acid, neat or indichloromethane, or HCl in dioxane or in ethyl acetate. The resultingammonium salt is then neutralized either prior to the coupling or insitu with basic solutions such as aqueous buffers, or tertiary amines indichloromethane or acetonitrile or dimethylformamide. When the Fmocgroup is used, the reagents of choice are piperidine or substitutedpiperidine in dimethylformamide, but any secondary amine can be used.The deprotection is carried out at a temperature between 0° C. and roomtemperature (rt or RT) usually 20-22° C.

Any of the amino acids having side chain functionalities must beprotected during the preparation of the peptide using any of theabove-described groups. Those skilled in the art will appreciate thatthe selection and use of appropriate protecting groups for these sidechain functionalities depend upon the amino acid and presence of otherprotecting groups in the peptide. The selection of such protectinggroups is important in that the group must not be removed during thedeprotection and coupling of the α-amino group.

For example, when Boc is used as the α-amino protecting group, thefollowing side chain protecting group are suitable:ρ-toluenesulfonyl(tosyl) moieties can be used to protect the amino sidechain of amino acids such as Lys and Arg; acetamidomethyl, benzyl (Bn),or tert-butylsulfonyl moieties can be used to protect the sulfidecontaining side chain of cysteine; bencyl (Bn) ethers can be used toprotect the hydroxy containing side chains of serine, threonine orhydroxyproline; and benzyl esters can be used to protect the carboxycontaining side chains of aspartic acid and glutamic acid.

When Fmoc is chosen for the α-amine protection, usually tert-butyl basedprotecting groups are acceptable. For instance, Boc can be used forlysine and arginine, tert-butyl ether for serine, threonine andhydroxyproline, and tert-butyl ester for aspartic acid and glutamicacid. Triphenylmethyl (Trityl) moiety can be used to protect the sulfidecontaining side chain of cysteine.

Once the elongation of the peptide is completed all of the protectinggroups are removed. When a liquid phase synthesis is used, theprotecting groups are removed in whatever manner is dictated by thechoice of protecting groups. These procedures are well known to thoseskilled in the art.

Further, the following guidance may be followed in the preparation ofcompounds of the present invention. For example, to form a compoundwhere R₄—C(O)—, R₄—S(O)₂, a protected P3 or the whole peptide or apeptide segment is coupled to an appropriate acyl chloride or sulfonylchloride respectively, that is either commercially available or forwhich the synthesis is well known in the art. In preparing a compoundwhere R₄O—C(O)—, a protected P3 or the whole peptide or a peptidesegment is coupled to an appropriate chloroformate that is eithercommercially available or for which the synthesis is well known in theart. For Boc-derivatives (Boc)₂O is used.

For example:

Cyclopentanol is treated with phosgene to furnish the correspondingchloroformate.

The chloroformate is treated with the desired NH₂-tripeptide in thepresence of a base such as triethylamine to afford thecyclopentylcarbamate.

In preparing a compound where R₄—N(R₅)—C(O)—, or R₄—NH—C(S)—, aprotected P3 or the whole peptide or a peptide segment is treated withphosgene followed by amine as described in Syn Lett. February 1995; (2);142-144 or is reacted with the commercially available isocyanate and asuitable base such as triethylamine.

In preparing a compound where R₄—N(R₅)—S(O₂), a protected P3 or thewhole peptide or a peptide segment is treated with either a freshlyprepared or commercially available sulfamyl chloride followed by amineas described in patent Ger. Offen. (1998), 84 pp. DE 19802350 or WO98/32748.

The α-carboxyl group of the C-terminal residue is usually protected asan ester (CPG) that can be cleaved to give the carboxylic acid.Protecting groups that can be used include: 1) alkyl esters such asmethyl, trimethylsilylethyl and t-butyl, 2) aralkyl esters such asbenzyl and substituted benzyl, or 3) esters that can be cleaved by mildbase treatment or mild reductive means such as trichloroethyl andphenacyl esters.

The resulting α-carboxylic acid (resulting from cleavage by mild acid,mild base treatment or mild reductive means) is coupled with aR₁SO₂NH₂[prepared by treatment of R₁SO₂Cl in ammonia saturatedtetrahydrofuran solution] in the presence of peptide coupling agent suchas CDI or EDAC in the presence of a base such as 4-dimethylaminopyridine(4-DMAP) and/or 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) to incorporatethe P1′ moiety, effectively assembling the tripeptide P1′-P1-P2-P3-APG.Typically, in this process, 1-5 equivalents of P1′ coupling agents areused.

Furthermore, if the P3 protecting group APG is removed and replaced witha B moiety by the methods described above, and the resultingα-carboxylic acid resulting from cleavage (resulting from cleavage bymild acid, mild base treatment or mild reductive means) is coupled witha R₁SO₂NH₂[prepared by treatment of R₁SO₂Cl in ammonia saturatedtetrahydrofuran solution or alternative methods described herein] in thepresence of peptide coupling agent such as CDI or EDAC in the presenceof a base such as 4-dimethylaminopyridine (4-DMAP) and/or1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) to incorporate the P1′ moiety,the tripeptide P1′-P1-P2-P3-B is prepared. Typically, in this process,1-5 equivalents of P1′ coupling agents are used.

Compounds of the present invention can be prepared by many methodsincluding those described in the examples, below, and as described inU.S. Pat. No. 6,323,180 and U.S. patent application Ser. No. 10/001,850filed on Nov. 20, 2001. The teachings of U.S. Pat. No. 6,323,180 andU.S. patent application Ser. No. 10/001,850 are incorporated herein, intheir entirety, by reference.

Scheme II further shows the general process wherein compounds of FormulaI are constructed by the coupling of tripeptide carboxylic acidintermediate (1) with a P1′ sulfonamide. (It should be noted that thegroups R₆, R₇, R₈, R₉, R₁₀, R₁₁ as shown below represent substituents ofthe heterocyclic system.) Said coupling reaction requires treatment ofcarboxylic acid (1) with a coupling reagent such as carbonyl diimidazolein a solvent such as THF, which can be heated to reflux, followed by theaddition of the formed derivative of (1), to the P1′ sulfonamide, in asolvent such as THF or methylene chloride in the presence of a base suchas DBU.

An alternative process for the construction of compounds of Formula I isshown in Scheme III. Therein the P1′ sulfonamide element is coupled tothe P1 element using the process employed in Scheme 1. The resultingP1-P1′ moiety can then be deprotected at it's amino terminus. In thisgeneral example a Boc protecting group is employed but one skilled inthe art would recognize that a number of suitable amino protectinggroups could be employed in this process. Said Boc protecting group canbe removed using acid such as trifluoroacetic acid in a solvent such asdichloroethane to provide the deprotected amine as the TFA salt. SaidTFA amine salt can be directly employed in the subsequent couplingreaction or as an alternative the TFA amine salt can be first convertedto the HCl amine salt, and this HCl amine salt is used in said couplingreaction as shown in Scheme III. The coupling of said HCl amine salt (3)with the carboxyl terminus a P4-P3-P2 intermediate can be achieved usingcoupling reagents, such as HATU, in solvents such as dichloromethane toprovide compounds of Formula I (4).

An alternative process for the construction of compounds of Formula I isshown in Scheme IV. Herein the hydrochloride salt of the P1-P1′ terminalamine (1) is coupled to the free carboxyl group of the P2 element usingcoupling agents such as PyBOP, in the presence of a base such asdiisopropyl amine, and in a solvent such as methylene chloride. Theresulting P2-P1-P1′ intermediate can be converted to compounds ofFormula I in a two step process wherein the first step is deprotectionof the P2 amine terminus using an acid such as TFA in a solvent such asmethylene chloride. The resulting trifluoroacetic acid salt can becoupled with the carboxyl terminus of the P4-P3 element using standardcoupling agents such as PyBop in the presence of base such asdiisopropyl amine, and using solvents such methylene chloride to providecompounds of Formula I (4).

The P4-P3-P2 intermediate utilized in the above schemes can beconstructed as previously described with a further description of thisprocess shown in general Scheme V. Therein the free carboxyl terminus ofthe P4-P3 intermediate (1), can be coupled to the amino terminus of theP2 element to provide the P4-P3-P2 dipeptide (2). The carboxyl terminusof the P4-P3-P2 intermediate can be deprotected by saponification of theester group to provide P4-P3-P2 as the free carboxylic acid (3).Intermediates like (3) can be converted to compounds of Formula I usingthe methods described herein.

Compounds of Formula I can also be converted into other compounds ofFormula I as described herein. An example of such a process is shown inScheme VI wherein a compound of Formula I (1) which bears a Boc group atthe P4 position is converted in a compound of Formula I (3) wherein saidcompound bears a urea group at the P4 position. The conversion of (1) to(3) can be carried out in a two step process the first of which is theconversion of (1) to amine (2) by treatment of (1) with an acid such asTFA in a solvent such as methylene chloride. The resulting amine TFAsalt can be treated with an isocyanate in the presence of one equivalentof base to provide a compound of Formula I (3) wherein the P3 moiety iscapped with a urea. As previously noted one skilled in the art willrecognize that intermediate (2) can be used as starting materials forthe preparation of compounds of Formula I wherein the P3 group is cappedwith an amide or a sulfonamide, or thiourea, or a sulfamide. Theconstruction of said compounds of Formula I can be achieved usingstandard conditions for the formation of said P4 functionalities fromamines.

In the construction of compounds of Formula I, the P1′ terminus isincorporated into the molecules using one of the general processesoutlined above and described in more detail below. In some examples theP1′ elements, that is the cycloalkyl- or alkyl sulfonamides arecommercially available or can be prepared from the corresponding alkyl-or cycloalkyl-sulfonyl chloride by treating said sulfonyl chloride withammonia. Alternatively, these sulfonamides can be synthesized using thegeneral process outline in Scheme VII. Therein commercially available3-chloro-propylsulfonyl chloride (1) is converted to a suitableprotected sulfonamide as for example by treatment with tert-butyl amine.The sulfonamide obtained (2) is then converted to the correspondingcycloalkylsulfonamide by treatment with two equivalents of a base suchas butyl lithium in a solvent such as THF at low temperature. Theresulting cycloalkylsulfonamide can be deprotected by treatment with anacid to provide the desired unprotected cycloalkylsulfoamide.

The P1 elements utilized in generating compounds of Formula I are insome cases commercially available, but are otherwise synthesized usingthe methods described herein and subsequently incorporated intocompounds of Formula I using the methods described herein. Thesubstituted P1 cyclopropylamino acids can be synthesized following thegeneral process outline in Scheme VIII.

Treatment of commercially available or easily synthesized imine (1) with1,4-dihalobutene (2) in presence of a base produces, provides theresulting imine (3). Acid hydrolysis of 3 then provides 4, which has anallyl substituent syn to the carboxyl group as a major product. Theamine moiety of 4 can protected using a Boc group to provide the fullyprotected amino acid 5. This intermediate is a racemate which can beresolved by an enzymatic process wherein the ester moiety of 5 iscleaved by a protease to provide the corresponding carboxylic acid.Without being bound to any particular theory, it is believed that thisreaction is selective in that one of the enantiomers undergoes thereaction at a much greater rate than its mirror image providing for akinetic resolution of the intermediate racemate. In the examples citedherein, the more preferred stereoisomer for integration into compoundsof Formula I is 5a which houses the (1R,2S) stereochemistry. In thepresence of the enzyme, this enantiomer does not undergo ester cleavageand thereby this enantiomer 5a is recovered from the reaction mixture.However, the less preferred enantiomer, 5b with houses the (1S,2R)stereochemistry undergoes ester cleavage, i.e., hydrolysis, to providethe free acid 6. Upon completion of this reaction, the ester 5a can beseparated from the acid product 6 by routine methods such as, forexample, aqueous extraction methods or chromotography.

Procedures for making P2 intermediates and compounds of Formula I areshown in the Schemes below. It should be noted that in many casesreactions are depicted for only one position of an intermediate.However, it is to be understood that such reactions could be used toimpart modifications to other positions within this intermediate.Moreover, said intermediates, reaction conditions and methods given inthe specific examples are broadly applicable to compounds with othersubstitution patterns. The general Schemes outlined below are followedwith examples herein. Both general and specific examples arenon-limiting, as for example the isoquinoline nucleus is shown as partof the general scheme, Scheme IX, however, this pathway represents aviable process for the construction of alternate heterocyclesubstituents as replacements for the isoquinoline element, such asquinolines, or pyridines.

Scheme IX shows the coupling of an N-protected C4-hydroxyproline moietywith a heterocycle to form intermediate (4) and the subsequentmodification of said intermediate (4) to a compound of Formula I by theprocess of peptide elongation as described herein. It should be notedthat in the first step, that is the coupling of the C4-hydroxy prolinegroup with the heteroaryl element, a base is employed. One skilled inthe art would recognized that this coupling can be done using bases suchas potassium tert-butoxide, or sodium hydride, in solvent such as DMF orDMSO or THF. This coupling to the isoquinoline ring system occurs at theCl position (numbering for isoquinoline ring system shown inintermediate 2 of Scheme IX) and is directed by the chloro group whichis displaced in this process. It should be noted that the alternativeleaving groups can be utilized at this position such as a fluoro asshown in the Scheme. Said fluoro intermediates (3) are available fromthe corresponding chloro compound using literature procedures describedherein. It should also be noted that the position of the leaving group(chloro or fluoro) in a given ring system can vary as shown in Scheme X,wherein the leaving group (fluoro in this example) is in the C6 positionof the isoquinoline ring system of intermediate (2).

It should be further noted that the position of the ring heteroatom(s)in intermediates like (2) of Scheme IX and Scheme X is also variable, asdefined by the term heterocycle described herein. In Scheme Xintermediate (2) can be coupled to a C4 hydroxy proline derivative toprovide the P2 element (3). This C6-substituted isoquinoline derivativecan be converted to compounds of Formula I using the methods describedherein.

An alternative to the method described above for the coupling of theC4-hydroxyproline to aromatics and heteroaromatics, is provided in theMitsunobu reaction as depicted in

step 1 of Scheme XI. In this general reaction Scheme a C4-hydroxyproline derivative is coupled to a quinazoline ring system. Thisreaction makes use of reagents such as triphenylphosphine and DEAD(diethylazodicarboxylate) in aprotic solvents such as THF or dioxane andcan be used for the formation of aryl and heteroaryl ethers. Note thatin the course of this coupling reaction the stereochemistry of the C4chiral center in the C4-hydroxyproline derivative is inverted andthereby it is necessary to use the C4-hydroxyproline derivative housingthe (S) stereochemistry at the C4 position as starting material. (asshown in Scheme XI). It should be noted that numerous modifications andimprovements of the Mitsunobu reaction have been described in theliterature, the teachings of which are incorporated herein.

In a subset of examples herein, isoquinolines are incorporated into thefinal compounds and specifically into the P2 region of said compounds.One skilled in the art would recognize that a number of general methodsare available for the synthesis of isoquinolines. Moreover, saidisoquinolines generated by these methods can be readily incorporatedinto final compounds of Formula I using the processes described herein.One general methodology for the synthesis of isoquinolines is shown inScheme XII, wherein cinnamic acid derivatives, shown in general form asstructure (2) are

-   Reference: N. Briet et al, Tetrahedron, 2002, 5761    converted to 1-chloroisoquinolines in a four step process. Said    chloroisoquinolines can be subsequently used in coupling reactions    to C4-hydroxyproline derivatives as described herein. The conversion    of cinnamic acids to chloroquinolines begins with the treatment of    cinnamic acid with an alkylcholorformate in the presence of a base.    The resulting anhydride is then treated with sodium azide which    results in the formation of an acylazide (3) as shown in the Scheme.    Alternate methods are available for the formation of acylazides from    carboxylic acids as for example said carboxylic acid can be treated    with diphenylphosphorylazide (DPPA) in an aprotic solvent such as    methylene chloride in the presence of a base. In the next step of    the reaction sequence the acylazide (3) is coverted to the    corresponding isoquinolone (4) as shown in the Scheme. In the event    the acylazide is heated to a temperature of approximately 190    degrees celcius in a high boiling solvent such a diphenylmethane.    This reaction is general and provides moderate to good yields of    substituted isoquinolone from the corresponding cinnamic acid    derivatives. It should noted that said cinnamic acid derivatives are    available commercially or can be obtained from the corresponding    benzaldehyde (1) derivative by direct condensation with malonic acid    or derivatives thereof and also by employing a Wittig reaction. The    intermediate isoquinolones (4) of Scheme XII can be converted to the    corresponding 1-chloroisoquinoline by treatment with phosphorous    oxychloride. This reaction is general and can be applied to any of    the isoquinolones, quinolones or additional heterocycles as shown    herein to covert a hydroxy substituent to the corresponding chloro    compound when said hydroxy is in conjugation with a nitrogen atom in    said heterocylic ring systems.

An alternative method for the synthesis of the isoquinoline ring systemis the Pomeranz-Fritsh procedure. This general method is outlined inScheme XIII. The process begins with the conversion of a benzaldehydederivative (1) to a functionalized imine (2). Said imine is thenconverted to the isoquinoline ring system by treatment with acid atelevated

temperature. This isoquinoline synthesis of Scheme XIII is general, andit should be noted that this process is particularly useful in procuringisoquinoline intermediates that are substituted at the C8 position(note: in intermediate (3) of Scheme XIII the C8 position of theisoquinoline ring is substituted with substituent R₁₁). The intermediateisoquinolines (3) can be converted to the corresponding1-chloroquinolines (5) in a two step process as shown. The first step inthis sequence is the formation of the isoquinoline N-oxide (4) bytreatment of isoquinoline (3) with meta-chloroperbenzoic acid in anaprotic solvent such as dichloromethane. Intermediate (4) can beconverted to the corresponding 1-chloroquinoline by treatment withphosphorous oxychloroide in refluxing chloroform. Note this two stepprocess is general and can be employed to procure chloroisoquinolinesand chloroquinolines from the corresponding isoquinolines and quinolinesrespectively.

Another method for the synthesis of the isoquinoline ring system isshown in Scheme XIV. In this process an ortho-alkylbenzamide derivative(1) is treated with a strong

base such as tert-butyl lithium in a solvent such as THF at lowtemperature. To this reaction mixture is then added a nitrilederivative, which undergoes an addition reaction with the anion derivedfrom deprotonation of (1), resulting in the formation of (2). Thisreaction is general and can be used for the formation of substitutedisoquinolines. Intermediate (2) of Scheme XIV can be converted to thecorresponding 1-chloroquinoline by the methods described herein.

An additional method for the synthesis of isoquinolines is shown inScheme XV. The deprotonation of intermediate (1) using tert-butyllithium is described above. In the present method however, saidintermediate anion is trapped by an ester, resulting in the formation ofintermediate (2) as shown below. In a subsequent reaction ketone (2) iscondensed with ammonium acetate at elevated temperature providing forthe formation of quinolone (3). This reaction is general and can beapplied for the construction of substituted isoquinolones which can thenbe converted to the corresponding 1-chloroisoquinolines as describedherein.

Yet an additional method for the construction of isoquinolines is foundin Scheme XVI. In the first step of this process an ortho-alkylaryliminederivatives such as (1) is subjected to deprotonation conditions(sec-butyl lithium, THF) and the resulting anion is quenched by

-   L. Flippin, J. Muchowski, JOC, 1993, 2631-2632

the addition of an activated carboxylic acid derivative such as aWeinreb amide. The resulting keto imine (2) can be converted to thecorresponding isoquinoline by condensation with ammonium acetate atelevated temperatures. This method is general and can be used for thesynthesis of substituted isoquinolines. Said isoquinolines can beconverted to the corresponding 1-chloroquinoline by the methodsdescribed herein.

The heterocycles described herein, and which are incorporated into thecompounds of Formula I can be further functionalized. It is obvious toone skilled in the art that additional functionalization of saidheterocycles can be done either before or after incorporation of thesefunctionalities into compounds of Formula I. The following Schemesillustrate this point. For example Scheme XVII shows the conversion of a1-chloro-

6-fluoro-isoquinoline to the corresponding1-chloro-6-alkoxy-isoquinoline species, by treatment of (1) of (eq. 1)with a sodium or potassium alkoxide species in the alcohol solvent fromwhich the alkoxide is derived at room temperature. In some cases it maybe necessary to heat the reaction to drive it to completion. Saidchloroquinoline can be incorporated into a compound of Formula I usingthe art described herein. Modifications of a P2 heterocyclic element canalso be done after it's incorporation into compounds of Formula I asshown in (eq. 2) of Scheme VXII. Specifically compounds such as (1) in(eq. 2) which contain a leaving group in the pthalazine nucleus can bedisplaced by a nucleophile such as an alkoxide in solvents such as thecorresponding alcohol from which the alkoxide is derived. These reactionscan be conducted at room temperature but in some cases it may benecessary to heat the reaction to drive it to completion.

Scheme XVIII provides a general example for the modification ofheterocycles as defined herein by employing palladium mediated couplingreactions. Said couplings can be employed to functionalize a heterocycleat each position of the ring system providing said ring is suitablyactivated or functionalized, as for example with a chloride as shown inthe Scheme. This sequence begins with 1-chloroisoquinoline (1) whichupon treatment with metachloroperbenzoic acid can be converted to thecorresponding N-oxide (2). Said intermediate (2) can be converted to thecorresponding 1,3-dichloroisoquinoline (3) by treatment with phosphorousoxychloride in refluxing chloroform. Intermediate (3) can be coupledwith N-Boc-4-hydroxyproline by the methods described herein to provideintermediate (5) as shown in the Scheme. Intermediate (5) can undergo aSuzuki coupling with an aryl boronic acid, in the presence of apalladium reagent and base, and in a solvent such as THF or toluene orDMF to provide the C3-arylisoquinoline intermediate (6).Heteroarylboronic acids can also be employed in this Pd mediatedcoupling process to provide C3-heteroarylisoquinolines. Intermediate (6)can be converted into final compounds of Formula I by the methodsdescribed herein.

Palladium mediated couplings of heteroaryl systems with aryl orheteroaryl elements can also be employed at a later synthetic stage inthe construction of compounds of Formula I as shown in Scheme IXX.Therein tripeptide acylsulfonamide intermediate (1) is coupled to a1-chloro-3-bromoisoquinoline (2) using the previously described processof alkoxide displacement of an heteroarylhalo moiety to provideintermediate (3). The coupling of (1) and (2) is most efficient in thepresence of a catalyst such as lanthanum chloride as described herein.The isoquinoline ring system of intermediate (3) can be furtherfunctionalized by employing either Suzuki couplings (Process 1:subjecting (3) to heteroaryl or aryl boronic acids in the presence of apalladium catalyst such as palladium tetra(triphenylphosphine) and abase such as cesium carbonate in solvents such as DMF) or Stillecouplings (Process 2: subjecting (3) to heteraryl or aryl tinderivatives in the presence of palladium catalyst such as palladiumtetra(triphenylphosphine in solvents such as toluene).

Palladium reactions can also be employed to couple C4-amino prolineelements with functionalized heterocycles. Scheme XX shows intermediate(1) coupling with a functionalized isoquinoline in the presence of apalladium catalyst and a base in a solvent such as toluene.Intermediates like (3) can be converted to compounds of Formula I usingthe methods described herein.

The construction of functionalized isoquinoline ring systems is alsopossible employing [4+2]cycloaddition reactions. For example (SchemeXXI) the use of vinyl isocyantes (1) in cycloaddition reactions withbenzyne precusors (2) provides functionalized isoquinolones (3). Saidisoquinolines can be incorporated into compounds of Formula I using themethods described herein.

The present invention also provides compositions comprising a compoundof the present invention, or a pharmaceutically acceptable salt, solvateor prodrug thereof, and a pharmaceutically acceptable carrier.Pharmaceutical compositions of the present invention comprise atherapeutically effective amount of a compound of the invention, or apharmaceutically acceptable salt, solvate or prodrug thereof, and apharmaceutically acceptable carrier, with a pharmaceutically acceptablecarrier, e.g., excipient, or vehicle diluent.

The active ingredient, i.e., compound, in such compositions typicallycomprises from 0.1 weight percent to 99.9 percent by weight of thecomposition, and often comprises from about 5 to 95 weight percent.

The pharmaceutical compositions of this invention may be administeredorally, parenterally or via an implanted reservoir. Oral administrationor administration by injection are preferred. In some cases, the pH ofthe formulation may be adjusted with pharmaceutically acceptable acids,bases or buffers to enhance the stability of the formulated compound orits delivery form. The term parenteral as used herein includessubcutaneous, intracutaneous, intravenous, intramuscular,intra-articular, intrasynovial, intrasternal, intrathecal, andintralesional injection or infusion techniques.

The pharmaceutical compositions may be in the form of a sterileinjectable preparation, for example, as a sterile injectable aqueous oroleaginous suspension. This suspension may be formulated according totechniques known in the art using suitable dispersing or wetting agentsand suspending agents. The details concerning the preparation of suchcompounds are known to those skilled in the art.

When orally administered, the pharmaceutical compositions of thisinvention may be administered in any orally acceptable dosage formincluding, but not limited to, capsules, tablets, and aqueoussuspensions and solutions. In the case of tablets for oral use, carrierswhich are commonly used include lactose and corn starch. Lubricatingagents, such as magnesium stearate, are also typically added. For oraladministration in a capsule form, useful diluents include lactose anddried corn starch. When aqueous suspensions are administered orally, theactive ingredient is combined with emulsifying and suspending agents. Ifdesired, certain sweetening and/or flavoring and/or coloring agents maybe added.

Other suitable carriers for the above noted compositions can be found instandard pharmaceutical texts, e.g. in “Remington's PharmaceuticalSciences”, 19 th ed., Mack Publishing Company, Easton, Pa., 1995.Further details concerning the design and preparation of suitabledelivery forms of the pharmaceutical compositions of the invention areknown to those skilled in the art.

Dosage levels of between about 0.01 and about 1000 milligram perkilogram (“mg/kg”) body weight per day, preferably between about 0.5 andabout 250 mg/kg body weight per day of the compounds of the inventionare typical in a monotherapy for the prevention and treatment of HCVmediated disease. Typically, the pharmaceutical compositions of thisinvention will be administered from about 1 to about 5 times per day oralternatively, as a continuous infusion. Such administration can be usedas a chronic or acute therapy. The amount of active ingredient that maybe combined with the carrier materials to produce a single dosage formwill vary depending upon the host treated and the particular mode ofadministration.

As the skilled artisan will appreciate, lower or higher doses than thoserecited above may be required. Specific dosage and treatment regimensfor any particular patient will depend upon a variety of factors,including the activity of the specific compound employed, the age, bodyweight, general health status, sex, diet, time of administration, rateof excretion, drug combination, the severity and course of theinfection, the patient's disposition to the infection and the judgmentof the treating physician. Generally, treatment is initiated with smalldosages substantially less than the optimum dose of the peptide.Thereafter, the dosage is increased by small increments until theoptimum effect under the circumstances is reached. In general, thecompound is most desirably administered at a concentration level thatwill generally afford antivirally effective results without causing anyharmful or deleterious side effects.

When the compositions of this invention comprise a combination of acompound of the invention and one or more additional therapeutic orprophylactic agent, both the compound and the additional agent areusually present at dosage levels of between about 10 to 100%, and morepreferably between about 10 and 80% of the dosage normally administeredin a monotherapy regimen.

When these compounds or their pharmaceutically acceptable salts,solvates or prodrugs are formulated together with a pharmaceuticallyacceptable carrier, the resulting composition may be administered invivo to mammals, such as man, to inhibit HCV NS3 protease or to treat orprevent HCV virus infection. Such treatment may also-be achieved usingthe compounds of this invention in combination with agents whichinclude, but are not limited to: immunomodulatory agents, such asinterferons; other antiviral agents such as ribavirin, amantadine; otherinhibitors of HCV NS3 protease; inhibitors of other targets in the HCVlife cycle such as helicase, polymerase, metalloprotease, or internalribosome entry site; or combinations thereof. The additional agents maybe combined with the compounds of this invention to create a singledosage form. Alternatively these additional agents may be separatelyadministered to a mammal as part of a multiple dosage form.

Accordingly, another aspect of this invention provides methods ofinhibiting HVC NS3 protease activity in patients by administering acompound of the present invention or a pharmaceutically acceptable saltor solvate thereof, wherein the substituents are as defined above.

In a preferred embodiment, these methods are useful in decreasing HCVNS3 protease activity in the patient. If the pharmaceutical compositioncomprises only a compound of this invention as the active component,such methods may additionally comprise the step of administering to saidpatient an agent selected from an immunomodulatory agent, an antiviralagent, a HCV protease inhibitor, or an inhibitor of other targets in theHCV life cycle such as, for example, helicase, polymerase, ormetalloprotease. Such additional agent may be administered to thepatient prior to, concurrently with, or following the administration ofthe compounds of this invention.

In an alternate preferred aspect, these methods are useful forinhibiting viral replication in a patient. Such methods can be useful intreating or preventing HCV disease.

The compounds of the invention may also be used as laboratory reagents.Compounds may be instrumental in providing research tools for designingof viral replication assays, validation of animal assay systems andstructural biology studies to further enhance knowledge of the HCVdisease mechanisms.

The compounds of this invention may also be used to treat or preventviral contamination of materials and therefore reduce the risk of viralinfection of laboratory or medical personnel or patients who come incontact with such materials, e.g., blood, tissue, surgical instrumentsand garments, laboratory instruments and garments, and blood collectionor transfusion apparatuses and materials.

EXAMPLES

The specific examples that follow illustrate the syntheses of thecompounds of the instant invention, and are not to be construed aslimiting the invention in sphere or scope. The methods may be adapted tovariations in order to produce compounds embraced by this invention butnot specifically disclosed. Further, variations of the methods toproduce the same compounds in somewhat different manner will also beevident to one skilled in the art.

Solution percentages express a weight to volume relationship, andsolution ratios express a volume to volume relationship, unless statedotherwise. Nuclear magnetic resonance (NMR) spectra were recorded eitheron a Bruker 300, 400 or 500 MHz spectrometer; the chemical shifts (δ)are reported in parts per million. Flash chromatography was carried outon silica gel (SiO₂) according to Still's flash chromatography technique(W. C. Still et al., J. Org. Chem., (1978), 43, 2923).

All Liquid Chromatography (LC) data were recorded on a Shimadzu LC-10ASliquid chromatograph using a SPD-10AV UV-Vis detector and MassSpectrometry (MS) data were determined with a Micromass Platform for LCin electrospray mode (ES+).

Unless otherwise noted, in the following examples each compound wasanalyzed by LC/MS, using one of seven methodologies, having thefollowing conditions.

Columns:

-   -   (Method A)—YMC ODS S7 C18 3.0×50 mm    -   (Method B)—YMC ODS-A S7 C18 3.0×50 mm    -   (Method C)—YMC S7 C18 3.0×50 mm    -   (Method D)—YMC Xterra ODS S7 3.0×50 mm    -   (Method E)—YMC Xterra ODS S7 3.0×50 mm    -   (Method F)—YMC ODS-A S7 C18 3.0×50 mm    -   (Method G)—YMC C18 S5 4.6×50 mm]        Gradient: 100% Solvent A/0% Solvent B to        0% Solvent A/100% Solvent B        Gradient time: 2 min. (A, B, D, F, G); 8 min. (C, E)        Hold time: 1 min. (A, B, D, F, G); 2 min. (C, E)        Flow rate: 5 mL/min        Detector Wavelength: 220 nm        Solvent A: 10% MeOH/90% H₂O/0.1% TFA        Solvent B: 10% H₂O/90% MeOH/0.1% TFA.

The abbreviations used in the present application, includingparticularly in the illustrative examples which follow, are well-knownto those skilled in the art. Some of the abbreviations used are asfollows:

-   rt room temperature-   Boc tert-butyloxycarbonyl-   DMSO dimethylsulfoxide-   EtOAc ethyl acetate-   t-BuOK potassium t-butoxide-   Et₂O diethyl ether-   TBME tert-butylmethyl ether-   THF tetrahydrofuran-   CDI carbonyldiimidazole-   DBU 1,8-diazabicyclo[5.4.0]undec-7-ene-   TFA trifluoroacetic acid-   NMM N-methylmorpholine-   HATU O-7-azabenzotriazol-1-yl-   HBTU O-{1H-benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium    hexafluorophosphate-   HOBT N-hydroxybenzotriazole-   PyBrop bromo-bis-pyrrolidine-phosphonium hexafluorophosphate-   DMF dimethylformamide-   MeOH methanol-   EDTA ethylenediaminetetraacetic acid-   HRMS high resolution mass spectrometry-   DMAP 4-dimethylaminopyridine-   DIPEA diisopropylethylamine-   DCM dichloromethane-   DCE dichloroethane

The compounds and chemical intermediates of the present invention,described in the following examples, were prepared according to thefollowing methods. It should be noted that the following exemplificationsection is presented in sections. The sections are titled Section Athrough K. Example numbers and compound numbers are not contiguousthroughout the entire Examples portion of the application and hence,each section indicates a “break” in the numbering. The numbering withineach section is generally contiguous. Section L describes the biologicalactivity of the compounds. Section M describes a subset of additionalcompounds that could be made using the methods described herein.

Section A Preparation of Intermediates Preparation of P1 Intermediates

The P1 intermediates described in this section can be used to preparecompounds of Formula I by the methods described herein.

I P1 Elements 1. Preparation ofracemic(1R,2S)/(1S,2R)-1-amino-2-vinylcyclopropane carboxylic acid ethylester

Method A Step 1

Glycine ethyl ester hydrochloride (303.8 g, 2.16 mole) was suspended intert-butylmethyl ether (1.6 L). Benzaldehyde (231 g, 2.16 mole) andanhydrous sodium sulfate (154.6 g, 1.09 mole) were added and the mixturecooled to 0° C. using an ice-water bath. Triethylamine (455 mL, 3.26mole) was added dropwise over 30 min and the mixture stirred for 48 h atrt. The reaction was then quenched by addition of ice-cold water (1 L)and the organic layer was separated. The aqueous phase was extractedwith tert-butylmethyl ether (0.5 L) and the combined organic phaseswashed with a mixture of saturated aqueous NaHCO₃ (1 L) and brine (1 L).The solution was dried over MgSO₄, concentrated in vacuo to afford 392.4g of the N-benzyl imine product as a thick yellow oil that was useddirectly in the next step. ¹H NMR (CDCl₃, 300 MHz) δ 1.32 (t, J=7.1 Hz,3H), 4.24 (q, J=7.1 Hz, 2H), 4.41 (d, J=1.1 Hz, 2H), 7.39-7.47 (m, 3H),7.78-7.81 (m, 2H), 8.31 (s, 1H).

Step 2

To a suspension of lithium tert-butoxide (84.06 g, 1.05 mol) in drytoluene (1.2 L), was added dropwise a mixture of the N-benzyl imine ofglycine ethyl ester (100.4 g, 0.526 mol) and trans-1,4-dibromo-2-butene(107.0 g, 0.500 mol) in dry toluene (0.6 L) over 60 min. Aftercompletion of the addition, the deep red mixture was quenched byaddition of water (1 L) and tert-butylmethyl ether (TBME, 1 L). Theaqueous phase was separated and extracted a second time with TBME (1 L).The organic phases were combined, 1 N HCl (1 L) was added and themixture stirred at room temperature for 2 h. The organic phase wasseparated and extracted with water (0.8 L). The aqueous phases were thencombined, saturated with salt (700 g), TBME (1 L) was added and themixture cooled to 0° C. The stirred mixture was then basified to pH 14by the dropwise addition of 10 N NaOH, the organic layer separated, andthe aqueous phase extracted with TBME (2×500 mL). The combined organicextracts were dried (MgSO₄) and concentrated to a volume of 1 L. To thissolution of free amine, was added BOC₂O or di-tert-butyldicarbonate(131.0 g, 0.6 mol) and the mixture stirred 4 days at rt. Additionaldi-tert-butyldicarbonate (50 g, 0.23 mol) was added to the reaction, themixture refluxed for 3 h, and was then allowed cool to room temperatureovernight. The reaction mixture was dried over MgSO₄ and concentrated invacuo to afford 80 g of crude material. This residue was purified byflash chromatography (2.5 Kg of SiO₂, eluted with 1% to 2% MeOH/CH₂Cl₂)to afford 57 g (53%) of racemicN-Boc-(1R,2S)/(1S,2R)-1-amino-2-vinylcyclopropane carboxylic acid ethylester as a yellow oil which solidified while sitting in therefrigerator: ¹H NMR (CDCl₃, 300 MHz) δ 1.26 (t, J=7.1 Hz, 3H), 1.46 (s,9H), 1.43-1.49 (m, 1H), 1.76-1.82 (br m, 1H), 2.14 (q, J=8.6 Hz, 1H),4.18 (q, J=7.2 Hz, 2H), 5.12 (dd J=10.3, 1.7 Hz, 1H), 5.25 (br s, 1H),5.29 (dd, J=17.6, 1.7 Hz, 1H), 5.77 (ddd, J=17.6, 10.3, 8.9 Hz, 1H); MSm/z 254.16 (M−1)

Step 3 Preparation of Racemic(1R,2S)/(1S,2R) 1-amino-2-vinylcyclopropanecarboxylic acid ethyl ester hydrochloride

N-Boc-(1R,2S)/(1S,2R)-1-amino-2-vinylcyclopropane carboxylic acid ethylester (9.39 g, 36.8 mmol) was dissolved in 4 N HCl/dioxane (90 ml, 360mmol) and was stirred for 2 h at rt. The reaction mixture wasconcentrated to supply (1R,2S)/(1S,2R)-1-amino-2-vinylcyclopropanecarboxylic acid ethyl ester hydrochloride in quanitative yield (7 g,100%). ¹H NMR (methanol-d₄) δ 1.32 (t, J=7.1, 3H), 1.72 (dd, J=10.2, 6.6Hz, 1H), 1.81 (dd, J=8.3, 6.6 Hz, 1H), 2.38 (q, J=8.3 Hz, 1H), 4.26-4.34(m, 2H), 5.24 (dd, 10.3, 1.3 Hz, 1H) 5.40 (d, J=17.2, 1H), 5.69-5.81 (m,1H).

Alternate Route for the Preparation of RacemicN-Boc-1-amino-2-vinylcyclopropane carboxylic acid ethyl esterhydrochloride

To a solution of potassium tert-butoxide (11.55 g, 102.9 mmol) in THF(450 mL) at −78° C. was added the commercially available N,N-dibenzylimine of glycine ethyl ester (25.0 g, 93.53 mmol) in THF (112 mL). Thereaction mixture was warmed to 0° C., stirred for 40 min, and was thencooled back to −78° C. To this solution was addedtrans-1,4-dibromo-2-butene (20.0 g, 93.50 mmol), the mixture stirred for1 h at 0° C. and was cooled back to −78° C. Potassium tert-butoxide(11.55 g, 102.9 mmol) was added, the mixture immediately warmed to 0°C., and was stirred one more hour before concentrating in vacuo. Thecrude product was taken up in Et₂O (530 mL), 1N aq. HCl solution (106mL, 106 mmol) added and the resulting biphasic mixture stirred for 3.5 hat rt. The layers were separated and the aqueous layer was washed withEt₂O (2×) and basified with a saturated aq. NaHCO₃ solution. The desiredamine was extracted with Et₂O (3×) and the combined organic extract waswashed with brine, dried (MgSO₄), and concentrated in vacuo to obtainthe free amine. This material was treated with a 4N HCl solution indioxane (100 mL, 400 mmol) and concentrated to afford(1R,2S)/(1S,2R)-1-amino-2-vinylcyclopropane carboxylic acid ethyl esterhydrochloride as a brown semisolid (5.3 g, 34% yield) identical to thematerial obtained from procedure A, except for the presence of a smallunidentified aromatic impurity (8%).

Resolution of N-Boc-(1R,2S)/(1S,2R)-1-amino-2-vinylcyclopropanecarboxylic acid ethyl ester

Resolution A

To an aqueous solution of sodium phosphate buffer (0.1 M, 4.25 liter(“L”), pH 8) housed in a 12 Liter jacked reactor, maintained at 39° C.,and stirred at 300 rpm was added 511 grams of Acalase 2.4 L (about 425mL) (Novozymes North America Inc.). When the temperature of the mixturereached 39° C., the pH was adjusted to 8.0 by the addition of a 50% NaOHin water. A solution of the racemicN-Boc-(1R,2S)/(1S,2R)-1-amino-2-vinylcyclopropane carboxylic acid ethylester (85 g) in 850 mL of DMSO was then added over a period of 40 min.The reaction temperature was then maintained at 40° C. for 24.5 h duringwhich time the pH of the mixture was adjusted to 8.0 at the 1.5 h and19.5 h time points using 50% NaOH in water. After 24.5 h, theenantio-excess of the ester was determined to be 97.2%, and the reactionwas cooled to room temperature (26° C.) and stirred overnight (16 h)after which the enantio-excess of the ester was determined to be 100%.The pH of the reaction mixture was then adjusted to 8.5 with 50% NaOHand the resulting mixture was extracted with MTBE (2×2 L). The combinedMTBE extract was then washed with 5% NaHCO₃ (3×100 mL), water (3×100mL), and evaporated in vacuo to give the enantiomerically pureN-Boc-(1R,2S)/-1-amino-2-vinylcyclopropane carboxylic acid ethyl esteras light yellow solid (42.55 g; purity: 97% @ 210 nm, containing noacid; 100% enantiomeric excess (“ee”).

The aqueous layer from the extraction process was then acidified to pH 2with 50% H₂SO₄ and extracted with MTBE (2×2 L). The MTBE extract waswashed with water (3×100 mL) and evaporated to give the acid as lightyellow solid (42.74 g; purity: 99% @ 210 nm, containing no ester).

ester acid High (+) ESI, C13H22NO4, (−) ESI, C11H16NO4, Resolution [M +H]⁺, cal. [M − H]⁻, Mass 256.1549, found cal. 226.1079, found Spec256.1542 226.1089 NMR observed chemical shift Solvent: CDCl₃ (proton δ7.24 ppm, C-13 δ 77.0 ppm) Bruker DRX-500C: proton 500.032 MHz, carbon125.746 MHz Proton (pattern) C-13 Proton (pattern) C-13 Position ppm ppmppm ppm  1 — 40.9 — 40.7  2 2.10 (q, J = 9.0 Hz) 34.1 2.17 (q, J = 9.0Hz) 35.0  3a 1.76 (br) 23.2 1.79 (br) 23.4  3b 1.46 (br) 1.51, (br)  4 —170.8 — 175.8  5 5.74 (ddd, J = 9.0, 133.7 5.75 (m) 133.4   10.0, 17.0Hz)  6a 5.25 (d, J = 17.0 Hz) 117.6 5.28 (d, J = 17.0 Hz) 118.1  6b 5.08(dd, J = 10.0, 5.12 (d, J = 10.5 Hz) 1.5 Hz)  7 — 155.8 — 156.2  8 —80.0 — 80.6  9 1.43 (s) 28.3 1.43 (s) 28.3 10 4.16 (m) 61.3 — — 11 1.23(t, J = 7.5 Hz) 14.2 — —Resolution B

To 0.5 mL 100 mM Heps.Na buffer (pH 8.5) in a well of a 24 well plate(capacity: 10 ml/well), 0.1 mL of Savinase 16.0 L (protease fromBacillus clausii) (Novozymes North America Inc.) and a solution of theracemic N-Boc-(1R,2S)/(1S,2R)-1-amino-2-vinylcyclopropane carboxylicacid ethyl ester (10 mg) in 0.1 mL of DMSO were added. The plate wassealed and incubated at 250 rpm at 40° C. After 18 h, enantio-excess ofthe ester was determined to be 44.3% as following: 0.1 mL of thereaction mixture was removed and mixed well with 1 mL ethanol; aftercentrifugation, 10 microliter (“μl”) of the supernatant was analyzedwith the chiral HPLC. To the remaining reaction mixture, 0.1 mL of DMSOwas added, and the plate was incubated for additional 3 days at 250 rpmat 40° C., after which four mL of ethanol was added to the well. Aftercentrifugation, 10 μl of the supernatant was analyzed with the chiralHPLC and enantio-excess of the ester was determined to be 100%.

Resolution C

To 0.5 ml 100 mM Heps.Na buffer (pH 8.5) in a well of a 24 well plate(capacity: 10 mL/well), 0.1 ml of Esperase 8.0 L, (protease fromBacillus halodurans) (Novozymes North America Inc.) and a solution ofthe racemic N-Boc-(1R,2S)/(1S,2R)-1-amino-2-vinylcyclopropane carboxylicacid ethyl ester (10 mg) in 0.1 mL of DMSO were added. The plate wassealed and incubated at 250 rpm at 40° C. After 18 hour, enantio-excessof the ester was determined to be 39.6% as following: 0.1 mL of thereaction mixture was removed and mixed well with 1 mL ethanol; aftercentrifugation, 10 μl of the supernatant was analyzed with the chiralHPLC. To the remaining reaction mixture, 0.1 mL of DMSO was added, andthe plate was incubated for additional 3 days at 250 rpm at 40° C.,after which four mL of ethanol was added to the well. Aftercentrifugation, 10 μl of the supernatant was analyzed with the chiralHPLC and enantio-excess of the ester was determined to be 100%.

Samples analysis was carried out in the following manner:

1) Sample preparation: About 0.5 ml of the reaction mixture was mixedwell with 10 volume of EtOH. After centrifugation, 10 μl of thesupernatant was injected onto HPLC column.

2) Conversion determination:

Column: YMC ODS A, 4.6×50 mm, S-5 μm

Solvent: A, 1 mM HCl in water; B, MeCN

Gradient: 30% B for 1 min; 30% to 45% B over 0.5 min; 45% B for 1.5 min;45% to 30% B over 0.5 min.

Flow rate: 2 ml/min

UV Detection: 210 nm

Retention time: acid, 1.2 min; ester, 2.8 min.

3) Enantio-excess determination for the ester:

Column: CHIRACEL OD-RH, 4.6×150 mm, S-5 μm

Mobile phase: MeCN/50 mM HClO₄ in water (67/33)

Flow rate: 0.75 ml/min.

UV Detection: 210 nm.

Retention time:

(1S,2R) isomer as acid: 5.2 min;

Racemate: 18.5 min and 20.0 min;

(1R,2S) isomer as ester: 18.5 min.

2. Preparation of N-Boc-(1R,2S)-1-amino-2-cyclopropylcyclopropanecarboxylic acid ethyl ester

A solution of N-Boc-(1R,2S)-1-amino-2-vinylcyclopropane carboxylic acid(255 mg, 1.0 mmol) in ether (10 mL) was treated with palladium acetate(5 mg, 0.022 mmol). The orange/red solution was placed under anatmosphere of N₂. An excess of diazomethane in ether was added dropwiseover the course of 1 h. The resulting solution was stirred at rt for 18h. The excess diazomethane was removed using a stream of nitrogen. Theresulting solution was concentrated by rotary evaporation to give thecrude product. Flash chromatography (10% EtOAc/hexane) provided 210 mg(78%) of N-Boc-(1R,2S)-1-amino-2-cyclopropylcyclopropane carboxylic acidethyl ester as a colorless oil. LC-MS (retention time: 2.13, similar tomethod A except: gradient time 3 min, Xterra MS C18 S7 3.0×50 mmcolumn), MS m/e 270 (M⁺+1).

3. 1-tert-butoxycarbonylamino-cyclopropane-carboxylic acid isCommercially Available

4. Preparation of 1-aminocyclobutanecarboxylic acid methylester•hydrochloride

1-aminocyclobutanecarboxylic acid (100 mg, 0.869 mmol)(Tocris) wasdissolved in 10 mL of MeOH, HCl gas was bubbled in for 2 h. The reactionmixture was stirred for 18 h, and then concentrated in vacuo to give 144mg of a yellow oil. Trituration with 10 mL of ether provided 100 mg ofthe titled product as a white solid. ¹H NMR (CDCl₃) δ 2.10-2.25 (m, 1H),2.28-2.42 (m, 1H), 2.64-2.82 (m, 4H), 3.87 (s, 3H), 9.21 (br s, 3H).

5. Preparation of racemic(1R,2R)/(1S,2S)1-Amino-2-ethylcyclopropanecarboxylic acid tert-butyl ester, Shown Below

Step 1: Preparation of 2-Ethylcyclopropane-1,1-dicarboxylic aciddi-tert-butyl ester, Shown Below

To a suspension of benzyltriethylammonium chloride (21.0 g, 92.2 mmol)in a 50% aqueous NaOH solution (92.4 g in 185 mL H₂O) was added1,2-dibromobutane (30.0 g, 138.9 mmol) and di-tert-butylmalonate (20.0g, 92.5 mmol). The reaction mixture was vigorously stirred 18 h at rt, amixture of ice and water was then added. The crude product was extractedwith CH₂Cl₂ (3×) and sequentially washed with water (3×), brine and theorganic extracts combined. The organic layer was dried (MgSO₄), filteredand concentrated in vacuo. The resulting residue was flashchromatographed (100 g SiO₂, 3% Et₂O in hexane) to afford the titledproduct (18.3 g, 67.8 mmol, 73% yield) which was used directly in thenext reaction.

Step 2: Preparation of racemic 2-Ethylcyclopropane-1,1-dicarboxylic acidtert-butyl ester, Shown Below

The product of Step 1 (18.3 g, 67.8 mmol) was added to a suspension ofpotassium tert-butoxide (33.55 g, 299.0 mmol) in dry ether (500 mL) at0° C., followed by H₂O (1.35 mL, 75.0 mmol) and was vigorously stirredovernight at rt. The reaction mixture was poured in a mixture of ice andwater and washed with ether (3×). The aqueous layer was acidified with a10% aq. citric acid solution at 0° C. and extracted with EtOAc (3×). Thecombined organic layers were washed with water (2×), brine, dried(MgSO₄) and concentrated in vacuo to afford the titled product as a paleyellow oil (10 g, 46.8 mmol, 69% yield).

Step 3: Preparation of (1R,2R)/(1S,2S)2-Ethyl-1-(2-trimethylsilanylethoxycarbonylamino)cyclopropane-carboxylicacid tert-butyl ester, Shown Below

To a suspension, of the product of Step 2 (10 g, 46.8 mmol) and 3 g offreshly activated 4 A molecular sieves in dry benzene (160 mL), wasadded Et₃N (7.50 mL, 53.8 mmol) and DPPA (11 mL, 10.21 mmol). Thereaction mixture was refluxed for 3.5 h, 2-trimethylsilyl-ethanol (13.5mL, 94.2 mmol) was then added, and the reaction mixture was refluxedovernight. The reaction mixture was filtered, diluted with Et₂O, washedwith a 10% aqueous citric acid solution, water, saturated aqueousNaHCO₃, water (2×), brine (2×), dried (MgSO₄) and concentrated in vacuo.The residue was suspended with 10 g of Aldrich polyisocyanate scavengerresin in 120 mL of CH₂Cl₂, stirred at rt overnight and filtered toafford the titled product (8 g, 24.3 mmol; 52%) as a pale yellow oil: ¹HNMR (CDCl₃) δ 0.03 (s, 9H), 0.97 (m, 5H), 1.20 (bm, 1H), 1.45 (s, 9H),1.40-1.70 (m, 4H), 4.16 (m, 2H), 5.30 (bs, 1H).

Step 4: Preparation of racemic(1R,2R)/(1S,2S)1-Amino-2-ethylcyclopropanecarboxylic acid tert-butyl ester, Shown Below

To the product of Step 3 (3 g, 9 mmol) was added a 1.0 M TBAF solutionin THF (9.3 mL, 9.3 mmol) and the mixture heated to reflux for 1.5 h,cooled to rt and then diluted with 500 ml of EtOAc. The solution wassuccessively washed with water (2×100 mL), brine (2×100 mL), dried(MgSO₄), concentrated in vacuo to provide the title intermediate

II P1′ Elements

The P1′ elements prepared below can be used to prepare compounds ofFormula I by using the methods described herein.

1. Preparation of cyclopropylsulfonamide

Step 1: Preparation of N-tert-Butyl-(3-chloro)propylsulfonamide

tert-Butylamine (3.0 mol, 315.3 mL) was dissolved in THF (2.5 L). Thesolution was cooled to −20° C. 3-Chloropropanesulfonyl chloride (1.5mol, 182.4 mL) was added slowly. The reaction mixture was allowed towarm to rt and stirred for 24 h. The mixture was filtered, and thefiltrate was concentrated in vacuo. The residue was dissolved in CH₂Cl₂(2.0 L). The resulting solution was washed with 1 N HCl (1.0 L), water(1.0 L), brine (1.0 L) and dried over Na₂SO₄. It was filtered andconcentrated in vacuo to give a slightly yellow solid, which wascrystallized from hexane to afford the product as a white solid (316.0g, 99%).

¹H NMR (CDCl₃) δ 1.38 (s, 9H), 2.30-2.27 (m, 2H), 3.22 (t, J=7.35 Hz,2H), 3.68 (t, J=6.2 Hz, 2H), 4.35 (b, 1H).

Step 2: Preparation of Cyclopropanesulfonic acid tert-butylamide

To a solution of N-tert-butyl-(3-chloro)propylsulfonamide (2.14 g, 10.0mmol) in THF (100 mL) was added n-BuLi (2.5 M in hexane, 8.0 mL, 20.0mmol) at −78° C. The reaction mixture was allowed to warm up to roomtemperature over period of 1 h. The volatiles were removed in vacuo. Theresidue was partitioned between EtOAC and water (200 mL, 200 mL). Theseparated organic phase was washed with brine, dried over Na₂SO₄,filtered and concentrated in vacuo. The residue was recrystallized fromhexane to yield the desired product as a white solid (1.0 g, 56%).

¹H NMR (CDCl₃) δ 0.98-1.00 (m, 2H), 1.18-1.19 (m, 2H), 1.39 (s, 9H),2.48-2.51 (m, 1H), 4.19 (b, 1H).

Step 3: Preparation of cyclopropylsulfonamide

A solution of cyclopropanesulfonic acid tert-butylamide (110.0 g, 0.62mol) in TFA (500 mL) was stirred at room temperature for 16 h. Thevolatile was removed in vacuo. The residue was recrystallized fromEtOAC/hexane (60 mL/240 mL) to yield the desired product as a whitesolid (68.5 g, 91%).

¹H NMR (DMSO-d₆) δ 0.84-0.88 (m, 2H), 0.95-0.98 (m, 2H), 2.41-2.58 (m,1H), 6.56 (b, 2H).

2. Alternate Procedure for the Preparation of Cyclopropyl Sulfonamide

To a solution of 100 mL of THF cooled to 0° C. was bubbled in gaseousammonia until saturation was reached. To this solution was added asolution of 5 g (28.45 mmol) of cyclopropylsulfonyl chloride (purchasedfrom Array Biopharma) in 50 mL of THF, the solution warmed to rtovernite and stirred one additional day. The mixture was concentrateduntil 1-2 mL of solvent remained, applied onto 30 g plug of SiO₂ (elutedwith 30% to 60% EtOAc/Hexanes) to afford 3.45 g (100%) of cyclopropylsulfonamide as a white solid. ¹H NMR (Methanol-d₄) δ 0.94-1.07 (m, 4H),2.52-2.60 (m, 1H); ¹³C NMR (methanol-d₄) δ 5.92, 33.01.

3. Preparation of Cyclobutyl Sulfonamide

To a solution of 5.0 g (37.0 mmol) of cyclobutyl bromide in 30 mL ofanhydrous diethyl ether (Et₂O) cooled to −78° C. was added 44 mL (74.8mmol) of 1.7M tert-butyl lithium in pentanes and the solution slowlywarmed to −35° C. over 1.5 h. This mixture was cannulated slowly into asolution of 5.0 g (37.0 mmol) freshly distilled sulfuryl chloride in 100mL of hexanes cooled to −40° C., warmed to 0° C. over 1 h and carefullyconcentrated in vacuo. This mixture was redissolved in Et₂O, washed oncewith some ice-cold water, dried (MgSO₄) and concentrated carefully. Thismixture was redissolved in 20 mL of THF, added dropwise to 500 mL ofsaturated NH₃ in THF and was allowed to stir overnite. The mixture wasconcentrated in vacuo to a crude yellow solid and was recrystallizedfrom the minimum amount of CH₂Cl₂ in hexanes with 1-2 drops of MeOH toafford 1.90 g (38%) of cyclobutylsulfonamide as a white solid. ¹H NMR(CDCl₃) δ 1.95-2.06 (m, 2H), 2.30-2.54 (m, 4H), 3.86 (p, J=8 Hz, 1H),4.75 (brs, 2H); ¹³C NMR (CDCl₃) δ 16.43, 23.93, 56.29. HRMS m/z (M−H)⁻calcd for C₄H₈NSO₂: 134.0276, found 134.0282.

4. Preparation of Cyclopentyl Sulfonamide

A solution of 18.5 mL (37.0 mmol) of 2M cyclopentyl-magnesium chloridein ether was added dropwise to a solution of 3.0 mL (37.0 mmol) freshlydistilled sulfuryl chloride (obtained from Aldrich) in 100 mL of hexanescooled to −78° C. The mixture was warmed to 0° C. over 1 h and was thencarefully concentrated in vacuo. This mixture was redissolved in Et₂O(200 mL), washed once with some ice-cold water (200 mL), dried (MgSO₄)and concentrated carefully. This mixture was redissolved in 35 mL ofTHF, added dropwise to 500 mL of saturated NH₃ in THF and was allowed tostir overnite. The mixture was concentrated in vacuo to a crude yellowsolid, the residue filtered through 50 g of silica gel using 70%EtOAc-hexanes as the eluent and the solution was then concentrated. Theresidue was recrystallized from the minimum amount of CH₂Cl₂ in hexaneswith 1-2 drops of MeOH to afford 2.49 g (41%) of cyclopentylsulfonamideas a white solid. ¹H NMR (CDCl₃) δ 1.58-1.72 (m, 2H), 1.74-1.88 (m, 2H),1.94-2.14 (m, 4H), 3.48-3.59 (m, 1H), 4.80 (bs, 2H); ¹³C NMR (CDCl₃) δ25.90, 28.33, 63.54; MS m/e 148 (M−H)⁻.

5. Preparation of Cyclohexyl Sulfonamide

A solution of 18.5 mL (37.0 mmol) of 2M cyclohexylmagnesium chloride(TCI Americas) in ether was added dropwise to a solution of 3.0 mL (37.0mmol) freshly distilled sulfuryl chloride in 100 mL of hexanes cooled to−78° C. The mixture was warmed to 0° C. over 1 h and was then carefullyconcentrated in vacuo. This mixture was redissolved in Et₂O (200 mL),washed once with some ice-cold water (200 mL), dried (MgSO₄) andconcentrated carefully This mixture was redissolved in 35 mL of THF,added dropwise to 500 mL of saturated NH₃ in THF and was allowed to stirovernite. The mixture was concentrated in vacuo to a crude yellow solid,the residue filtered through 50 g of silica gel using 70% EtOAc-hexanesas the eluent and was concentrated. The residue was recrystallized fromthe minimum amount of CH₂Cl₂ in hexanes with 1-2 drops of MeOH to afford1.66 g (30%) of cyclohexyl-sulfonamide as a white solid: ¹H NMR (CDCl₃)δ 1.11-1.37 (m, 3H), 1.43-1.56 (m, 2H), 1.67-1.76 (m, 1H), 1.86-1.96 (m,2H), 2.18-2.28 (m, 2H), 2.91 (tt, J=12, 3.5 Hz, 1H), 4.70 (bs, 2H); ¹³CH NMR (CDCl₃) δ 25.04, 25.04, 26.56, 62.74; MS m/e 162 (M−1).

6. Preparation of Neopentylsulfonamide

Following the procedure for the prep of cyclohexyl sulfonamide, 49 mL(37 mmol) of 0.75M neopentylmagnesium chloride (Alfa) in ether wasconverted to 1.52 g (27%) of neopentylsulfonamide as a white solid. ¹HNMR (CDCl₃) δ 1.17 (s, 9H), 3.12 (s, 2H), 4.74 (brs, 2H); ¹³C NMR(CDCl₃) δ 29.46, 31.51, 67.38; MS m/e 150 (M−1)⁻.

7. Preparation of cyclobutylcarbonyl-sulfonamide

A solution of 12.3 g (83 mmol) of cyclobutylcarbonyl bromide (Aldrich)and 13.7 g (91 mmol) of sodium iodide in 150 mL of acetone was refluxedovernite and then cooled to rt. The inorganic solids were filtered offand the acetone and cyclopropylcarbonyl iodide (8.41 g, 46%) distilledoff at ambient and 150 torr at 80° C., respectively.

A solution of 4.0 g (21.98 mmol) of cyclobutyl carbonyl iodide in 30 mLof anhydrous diethyl ether (Et₂O) cooled to −78° C. was cannulated intoa solution of 17 mL (21.98 mmol) of 1.3M sec-butyl lithium incyclohexanes and the solution was stirred for 5 min. To this mixture wascannulated a solution of 3.0 g (21.98 mmol) of freshly distilledsulfuryl chloride in 110 mL of hexanes cooled to −78° C., the mixturewarmed to rt over 1 h and was then carefully concentrated in vacuo. Thismixture was redissolved in Et₂O, washed once with some ice-cold water,dried (MgSO₄) and concentrated carefully. This mixture was redissolvedin 30 mL of THF, added dropwise to 500 mL of saturated NH₃ in THF andwas allowed to stir overnite. The mixture was concentrated in vacuo to acrude yellow solid and was recrystallized from the minimum amount ofCH₂Cl₂ in hexanes with 1-2 drops of MeOH to afford 1.39 g (42%) ofcyclobutyl carbonylsulfonamide as a white solid. ¹H NMR (CDCl₃) δ1.81-2.03 (m, 4H), 2.14-2.28 (m, 2H), 2.81-2.92 (m, 1H), 3.22 (d, J=7Hz, 2H), 4.74 (brs, 2H); ¹³C NMR (CDCl₃) δ 19.10, 28.21, 30.64, 60.93;MS m/e 148 (M−1)⁻. time: 1.73, method B), 818 (M⁺+H)

8. Preparation of cyclopropylcarbonyl-sulfonamide

Using the procedure employed for the preparation ofcyclobutylcarbonyl-sulfonamide, cyclopropylcarbonyl sulfonamide wasprepared from cyclopropylcarbonyl bromide (Aldrich) (see also JACS 1981,p. 442-445). ¹H NMR (CDCl₃) δ 0.39-0.44 (m, 2H), 0.67-0.76 (m, 2H),1.13-1.27 (m, 1H), 3.03 (d, J=7.3 Hz, 2H), 4.74 (brs, 2H); ¹³C NMR(CDCl₃) δ 4.33, 5.61, 59.93; MS m/e 134 (M−1).

III Heterocycles to be Used as Starting Material in the Construction ofP2 Elements for Subsequent Incorporation into Compounds of Formula I 1.Isoquinolines

Isoquinoline (1) and substituted analogues thereof, can be incorporatedinto P2 elements using the two methods outline above and described indetail herein. Said P2 elements (3) can then be converted into compoundsof Formula I using procedures analogous to those described herein forsimilar isoquinoline analogues.

2. Isoxazolepyridine and Oxazolepyridine (1)

Isoxazole and oxazole heterocycle (1) and analogues thereof can beprepared using know chemistry and incorporated into compounds of FormulaI using the chemistry described herein for similar isoxazolepyridineintermediates as shown in section B.

Section B

In Section B the following conditions were used for LC/MS analysis.

Columns:

-   -   Method A: YMC ODS-A C18 S7 (4.6×33 mm)    -   Method B: YMC Xterra ODS S7 (3.0×50 mm)    -   Method C: Xterra ms C18 (4.6×33 mm)    -   Method D: YMC ODS-A C18 S3 (4.6×33 mm)        Gradient: 100% solvent A/0% solvent B to 0% solvent A/100%        solvent B        Gradient time: 3 min.        Hold Time: 1 min.        Flow Rate: 5 mL/min.        Detector Wavelength: 220 nm.        Solvents: Solvent A: 10% MeOH/90% water/0.1% TFA. Solvent B: 90%        MeOH/10% water/0.1% TFA.

The following conditions were used for prep-HPLC separation.

Columns: Phenomenex-Luna 30×100 mm, S5

Gradient: 60% solvent A/10% solvent B to 0% solvent A/100% solvent B

Gradient time: 15 min.

Stop Time: 20 min.

Flow Rate: 30 mL/min.

Detector Wavelength: 220 nm.

Solvents: Solvent A: 10% MeOH/90% water/0.1% TFA. Solvent B: 90%MeOH/10% water/0.1% TFA.

Example 1 Preparation of Compound 1

Step 1:

A mixture of 3,5-dimethyl-4-nitro-isoxazole (1.42 g, 10.0 mmol),phenylacetaldehyde (1.32 g, 11.0 mmol) in piperidine (1 mL) and ethanol(10 mL) was heated to reflux for 16 h. After cooling down to the ambienttemperature, the product precipitated out was collected by filtration.The cake was washed with cold ethanol thoroughly to afford 1.20 g (53%)of the desired product as a white solid.

¹H NMR (CDCl₃) δ 2.87 (s, 3H), 7.46-7.50 (m, 3H), 7.56 (d, J=8.5 Hz,1H), 7.7-7.80 (m, 2H);

LC-MS (retention time: 1.19 min, method B), MS m/z 227 (M⁺+H).

Step 2:

A solution of 3-methyl-5-phenyl-isoxazolo[4,5-b]pyridine 4-oxide (1.00g, 4.40 mmol) and POCl₃ (2.71 g, 17.7 mmol) in chloroform (10 mL) washeated to reflux for 1 h. After cooling down to the ambient temperature,the final solution was diluted with chloroform (50 mL) and washed withNaHCO₃ (aq.) (two 50 mL portions) and brine, dried over MgSO₄, filtered,evaporated. The residue was purified by flash chromatography (4:1hexane-EtOAc) to afford 790 mg (73%) of the desired product as a whitesolid.

¹H NMR (CDCl₃) δ 2.72 (s, 3H), 7.46-7.54 (m, 3H), 7.91 (s, 1H),8.00-8.03 (m, 2H);

LC-MS (retention time: 1.76 min, method B), MS m/z 245, 247 (M⁺+H).

Step 3:

To a mixture of 4-hydroxy-pyrrolidine-2-carboxylic acid methyl ester(H-Hyp-OMe HCl) (1.81 g, 10.0 mmol), HATU (5.70 g, 15.0 mmol), andN—BOC-t-butyl-L-glycine (2.42 g, 10.5 mmol) in CH₂Cl₂ (100 mL) was addedDIPEA (3.47 g, 31.0 mmol) at 0° C. After stirring at the ambienttemperature for 12 h, the formed solution was diluted with CH₂Cl₂ (100mL), washed with iced 5% citric acid (aq). The organic layer was washedwith 5% citric acid, 1M NaOH, brine respectively, dried over MgSO₄, andfiltered. The filtrate was evaporated in vacuo to provide 3.55 g (99%)of the desired product as an off-white foam. This product was used forthe next reaction as crude without further purification.

¹H NMR (CD₃OD) δ 1.04 (s, 9H), 1.43 (s, 9H), 1.99-2.03 (m, 1H),2.20-2.30 (m, 1H), 3.69 (s, 3H), 3.70-3.79 (m, 2H), 4.28 (b, 1H), 4.46(b, 1H), 4.74-4.80 (m, 1H);

LC-MS (retention time: 1.28 min, method B), MS m/z 359 (M⁺+H).

Step 4:

A mixture of the product of Step 3 (3.55 g, 9.9 mmol) in THF (50 mL),MeOH (50 mL) and LiOH monohydrate (0.83 g, 19.9 mmol in 50 mL H₂O) wasstirred at the ambient temperature over night. After removal of thevolatiles in vacuo, the residue was dissolved in 0.1 M NaOH (100 mL).This aqueous solution was washed with ether (50 mL), acidified by 1M HClto pH4. Extracted with EtOAc (100 mL). The organic layer was washed with5% citric acid and brine, dried over MgSO₄, evaporated to dryness togive 3.20 g (95%) of the desired product as a white foam.

This product was used as crude without further purification.

¹H NMR (CD₃OD) δ 1.02 (s, 9H), 1.43 (s, 9H), 2.01-2.09 (m, 1H),2.25-2.32 (m, 1H), 3.70-3.85 (m, 2H), 4.26-4.30 (m, 1H), 4.46-4.51 (m,2H), 6.37-6.41 (m, 1H);

LC-MS (retention time: 1.14 min, method B), MS m/z 345 (M⁺+H).

Step 5:

To a solution of the product of Step 4 (1.01 g, 2.93 mmol) in DMSO (30mL) was added potassium tert-butoxide (1.02 g, 9.08 mmol). The formedsolution was stirred at the ambient temperature for 1 h before additionof 7-chloro-3-methyl-5-phenyl-isoxazolo[4,5-b]pyridine (0.75 g, 3.08mmol). The final solution was stirred for 12 h. Then was quenched withiced water, acidified with 1M HCl to pH 4, extracted with EtOAc (two 200mL portions). The organic layers were washed with brine, dried overMgSO₄, filtered, evaporated. The residue was purified by prep-HPLC (60%B-100% B, 15 min gradient) to afford 305 mg (19%) of the desired productas a pale yellow solid.

¹H NMR (CD₃OD) δ 1.02 (s, 9H), 1.17 (s, 9H), 2.37-2.47 (m, 1H), 2.64 (s,3H), 2.85-2.93 (m, 1H), 4.00-4.08 (m, 1H), 4.14 (b, 1H), 4.49-4.55 (m,1H), 4.62-4.71 (m, 1H), 5.70 (m, 1H), 7.45-7.53 (m, 3H), 7.56 (s, 1H),8.03-8.06 (m, 2H);

LC-MS (retention time: 1.89 min, method B), MS m/z 553 (M⁺+H).

Step 6a

As described in section A. Step 6b:

To a solution of1(R)-tert-butoxycarbonylamino-2(S)-vinyl-cyclopropanecarboxylic acidethyl ester, the product of Step 6a (3.28 g, 13.2 mmol) in THF (7 mL)and methanol (7 mL) was added a suspension of LiOH (1.27 g, 53.0 mmol)in water (14 mL). The mixture was stirred overnight at room temperatureand quenched with 1N NaOH (15 mL) and water (20 mL). The resultingmixture was washed with EtOAc (20 mL), and the organic phase wasextracted with 20 mL 0.5N NaOH. The combined aqueous phases wereacidified with 1N HCl until pH 4 and extracted with EtOAc (3×40 mL). Thecombined organic extracts were washed with brine and dried (MgSO₄) toyield the title compound as a white solid (2.62 g, 87%).

¹H NMR: (DMSO-d₆) δ 1.22-1.26 (m, 1H), 1.37 (s, 9H), 1.50-1.52 (m, 1H),2.05 (q, J=9 Hz, 1H), 5.04 (d, J=10 Hz, 1H), 5.22 (d, J=17 Hz, 1H),5.64-5.71 (m, 1H), 7.18, 7.53 (s, NH (rotamers), 12.4 (br s, 1H));

LC-MS (retention time: 1.67 min, method B), MS m/z 228 (M⁺+H).

Step 7:

A solution of the product of Step 6 (2.62 g, 11.5 mmol) and CDI (2.43 g,15.0 mmol) in THF (40 mL) was heated at reflux for 50 min undernitrogen. The solution was cooled to room temperature and transferred bycannula to a solution of cyclopropylsulfonamide (1.82 g, 15.0 mmol) inTHF (10 mL). To the resulting solution was added DBU (2.40 mL, 16.1mmol) and stirring was continued for 20 h. The mixture was quenched with1N HCl to pH 1 and THF was evaporated in vacuo. The suspension wasextracted with EtOAc (2×50 mL) and the combined organic extracts dried(Na₂SO₄). Purification by recystallization from hexanes-EtOAc (1:1)afforded the title compound (2.4 g) as a white solid. The mother liquorwas purified by a Biotage 40S column (eluted 9% acetone in DCM) to givea second batch of the title compound (1.1 g). Both batches were combined(total yield 92%).

¹H NMR: (DMSO-d₆) δ 0.96-1.10 (m, 4H), 1.22 (dd, J=5.5, 9.5 Hz, 1H),1.39 (s, 9H), 1.70 (t, J=5.5 Hz, 1H), 2.19-2.24 (m, 1H), 2.90 (m, 1H),5.08 (d, J=10 Hz, 1H), 5.23 (d, J=17 Hz, 1H), 5.45 (m, 1H), 6.85, 7.22(s, NH (rotamers);

LC-MS (retention time: 1.70 min, method B), MS m/z 331 (M⁺+H).

Step 8:

A solution of the product of Step 7 (3.5 g, 10.6 mmol) in DCM (35 mL)and TFA (32 mL) was stirred at room temperature for 1.5 h. The volatileswere removed in vacuo and the residue suspended in 1N HCl in diethylether (20 mL) and concentrated in vacuo. This procedure was repeatedonce. The resulting mixture was triturated from pentane and filtered togive the title compound as a hygroscopic, off-white solid (2.60 g, 92%).

¹H NMR: (DMSO-d₆) δ 1.01-1.15 (m, 4H), 1.69-1.73 (m, 1H), 1.99-2.02 (m,1H), 2.38 (q, J=9 Hz, 1H), 2.92-2.97 (m, 1H), 5.20 (d, J=1 Hz, 1H), 5.33(d, J=17 Hz, 1H), 5.52-5.59 (m, 1H), 9.17 (br s, 3H);

LC-MS (retention time: 0.24 min, method B), MS m/z 231 (M⁺+H).

Step 9:

To an iced mixture of the product of Step 5 (70 mg, 0.13 mmol),(1R,2S)-cyclopropanesulfonic acid(1-amino-2-vinyl-cyclopropanecarbonyl)amide hydrochloride, the productof Step 8 (37 mg, 0.14 mmol) and HATU (72 mg, 0.19 mmol) in DCM (2 mL)was added diisopropylethylamine (50 mg, 0.39 mmol). The formed solutionwas allowed to warm up to the ambient temperature for 12 h andevaporated in vacuo. The residue was purified by prep-HPLC (60% B-100%B, 15 min gradient) to afford 52 mg (54%) of Compound 1 as a grayishsolid.

¹H NMR (CD₃OD) δ 0.96-1.09 (m, 12H), 1.16-1.25 (m, 10H), 1.44-1.48 (m,1H), 1.87-1.91 (m, 1H), 2.20-2.40 (m, 2H), 2.63-2.65 (m, 4H), 2.89-2.98(m, 1H), 4.08-4.20 (m, 2H), 4.44-4.65 (m, 2H), 5.13 (d, J=11.7 Hz, 1H),5.32 (d, J=15 Hz, 1H), 5.72-5.85 (b, 2H), 6.62 (d, J=15.0 Hz, 1H),7.46-7.53 (m, 3H), 7.58 (s, 1H), 8.04-8.07 (m, 2H);

LC-MS (retention time: 1.92 min, method B), MS m/z 765 (M⁺+H).

Example 2 Preparation of Compound 2

Step 1:

A mixture of 2-amino-6-methylpyridine (1.08 g, 10.0 mmol), ethylbenzoylacetate (2.30 g, 12.0 mmol) and polyphosphoric acid (6.00 g, 61.2mmol) was heated to 110° C. for 5 h. After cooling to the ambienttemperature, the mixture was poured into iced water (20 mL) andneutralized to pH 7 with 10 M NaOH. Extracted with CHCl₃. The organiclayer was washed with brine, dried over MgSO₄, filtered, evaporated. Theresidue was purified by flash chromatography (1:1 hexane-EtOAc) toafford 510 mg (22%) of the desired product as a pale yellow solid.

¹H NMR (CDCl₃) δ 3.08 (s, 3H), 6.64 (d, J=7.0 Hz, 1H), 6.71 (s, 1H),7.42-7.52 (m, 5H), 8.04-8.06 (m, 2H);

LC-MS (retention time: 1.21 min, method B), MS m/z 237 (M⁺+H).

Step 2:

A solution of 6-methyl-2-phenyl-pyrido[1,2a]pyrimidin-4-one (489 mg,2.07 mmol) in melted diphenyl ether (5 mL) was heated to gentle refluxfor 5 h. After cooling to the ambient temperature, the formed suspensionwas diluted with diethyl ether (10 mL), filtered. The cake was washedwith diethyl ether thoroughly to afford 450 mg (92%) of the desiredproduct as a brownish solid.

LC-MS (retention time: 1.25 min, method B), MS m/z 237 (M⁺+H).

Step 3:

A suspension of 7-methyl-2-phenyl-1H-[1,8]naphthyridin-4-one (450 mg,1.91 mmol) in POCl₃ (10 mL) was heated to gentle reflux for 3 h.Evaporated in vacuo. The residue was poured into iced water (20 mL) andneutralized to pH 10 with 10 M NaOH. Extracted with CHCl₃. The organiclayer was washed with brine, dried over MgSO₄, filtered, evaporated. Theresidue was purified by flash chromatography (2:1 hexane-EtOAc) toafford 450 mg (92%) of the desired product as a pink solid.

¹H NMR (CD₃OD) δ 2.80 (s, 3H), 7.54-7.56 (m, 3H), 7.61 (d, J=8.4 Hz,1H), 8.25-8.30 (m, 3H), 8.58 (d, J=8.4 Hz, 1H);

LC-MS (retention time: 1.39 min, method B), MS m/z 255, 257 (M⁺+H).

Step 4:

This product was prepared by the same procedure as described in Example1, Step 5, except using 4-chloro-7-methyl-2-phenyl-[1,8]naphthyridinefrom Example 2, Step 3 instead.

LC-MS (retention time: 1.55 min, method B), MS m/z 563 (M⁺+H).

Step 5:

Compound 2 was prepared by the same procedure as described in Example 1,Step 9, except using the product of Example 2, Step 4 instead.

¹H NMR (CD₃OD) δ 1.01-1.10 (m, 12H), 1.21-1.26 (m, 10H), 1.40-1.45 (m,1H), 1.86-1.91 (m, 1H), 2.20-2.29 (m, 1H), 2.39-2.49 (m, 1H), 2.72-2.81(m, 1H), 2.92-2.95 (m, 4H), 4.10-4.16 (m, 2H), 4.55-4.65 (m, 2H), 5.14(d, J=12.0 Hz, 1H), 5.30 (d, J=15.0 Hz, 1H), 5.67-5.82 (m, 2H),7.60-7.80 (m, 3H), 7.78 (d, J=8.6 Hz, 1H), 7.87 (s, 1H), 8.26-8.29 (m,2H), 8.95 (d, J=8.4 Hz, 1H);

LC-MS (retention time: 1.62 min, method B), MS m/z 775 (M⁺+H).

Example 3 Preparation of Compound 3

Step 1:

To a solution of 4-methoxyphenethyl alcohol (1.52 g, 10.0 mmol) inCH₂Cl₂ (50 mL) at 0° C. was added Dess-Martin reagent (4.45 g, 10.5mmol) in one portion.

The formed mixture was allowed to warm to the ambient temperature for 1h. Washed with sat. Na₂S₂O₃ (aq) and 1M NaOH, brine respectively. Driedover MgSO₄, evaporated in vacuo to give 1.50 g (100%) of the desiredaldehyde as a viscous oil. This product was used as crude without anyfurther purification.

Step 2:

A solution of 3,5-dimethyl-4-nitro-isoxazole (142 mg, 1.0 mmol),4-methoxy-phenylacetaldehyde from Example 3, Step 1 (180 mg, 1.1 mmol)in piperidine (0.1 mL) and ethanol (2 mL) was heated to reflux for 12 h.After cooling down to the ambient temperature, the product precipitatedout was collected by filtration. The cake was washed with cold ethanolthoroughly to afford 130 mg (51%) of the desired product as a grayishsolid.

¹H NMR (CDCl₃) δ 2.88 (s, 3H), 3.87 (s, 3H), 7.02 (d, J=8.5 Hz, 2H),7.50 (d, J=9.0 Hz, 1H), 7.57 (d, J=9.0 Hz, 1H), 7.81 (d, J=8.5 Hz, 2H);

LC-MS (retention time: 1.24 min, method B), MS m/z 257 (M⁺+H).

Step 3:

This product was prepared by the same procedure as described in Example1, Step 2, except using the product of Example 3, Step 2 instead.

¹H NMR (CDCl₃) δ 2.70 (s, 3H), 3.87 (s, 3H), 7.00-7.03 (m, 2H), 7.84 (s,1H), 7.96-7.98 (m, 2H);

LC-MS (retention time: 1.96 min, method B), MS m/z 275, 277 (M⁺+H).

Step 4:

This product was prepared by the same procedure as described in Example1, Step 5, except using the product of Example 3, Step 3 instead.

¹H NMR (CD₃OD) δ 1.02 (s, 9H), 1.18 (s, 9H), 2.39-2.43 (m, 1H), 2.63 (s,3H), 2.75-2.80 (m, 1H), 3.87 (s, 3H), 4.00-4.08 (m, 1H), 4.17 (b, 1H),4.49-4.55 (m, 1H), 4.62-4.71 (m, 1H), 5.68 (b, 1H), 7.05 (d, J=8.5 Hz,2H), 7.49 (s, 1H), 8.00 (d, J=8.5 Hz, 2H);

LC-MS (retention time: 1.89 min, method B), MS m/z 583 (M⁺+H).

Step 5:

Compound 3 was prepared by the same procedure as described in Example 1,Step 9, except using the product of Example 3, Step 4 instead.

¹H NMR (CD₃OD) δ 1.01-1.09 (m, 12H), 1.17-1.26 (m, 10H), 1.44-1.47 (m,1H), 1.87-1.91 (m, 1H), 2.20-2.40 (m, 2H), 2.63-2.65 (m, 4H), 2.89-2.98(m, 1H), 3.87 (s, 3H), 4.08-4.20 (m, 2H), 4.44-4.65 (m, 2H), 5.13 (d,J=11.7 Hz, 1H), 5.32 (d, J=15.0 Hz, 1H), 5.72-5.85 (m, 2H), 7.05 (d,J=8.5 Hz, 2H), 7.06 (s, 1H), 8.01 (d, J=8.5 Hz, 2H);

LC-MS (retention time: 1.96 min, method B), MS m/z 795 (M⁺+H).

Example 4 Preparation of Compound 4

Step 1:

This product was prepared by the same procedure as described in Example3, Step 1&2, except using 4-fluorophenethyl alcohol instead.

LC-MS (retention time: 1.18 min, method B), MS m/z 245 (M⁺+H).

Step 2:

This product was prepared by the same procedure as described in Example1, Step 2, except using the product of Example 4, Step 1 instead.

¹H NMR (CDCl₃) δ 2.71 (s, 3H), 7.17-7.20 (m, 2H), 7.86 (s, 1H),8.00-8.02 (m, 2H);

LC-MS (retention time: 1.71 min, method B), MS m/z 263, 265 (M⁺+H).

Step 3:

This product was prepared by the same procedure as described in Example1, Step 5, except using the product of Example 4, Step 2 instead.

LC-MS (retention time: 1.91 min, method B), MS m/z 571 (M⁺+H).

Step 4:

Compound 4 was prepared by the same procedure as described in Example 1,Step 9, except using the product of Example 4, Step 3 instead.

¹H NMR (CD₃OD) δ 1.01-1.09 (m, 12H), 1.17-1.26 (m, 10H), 1.44-1.47 (m,1H), 1.87-1.91 (m, 1H), 2.20-2.40 (m, 2H), 2.63-2.65 (m, 4H), 2.89-2.98(m, 1H), 4.08-4.20 (m, 2H), 4.44-4.65 (m, 2H), 5.13 (d, J=11.7 Hz, 1H),5.32 (d, J=15.0 Hz, 1H), 5.72-5.85 (m, 2H), 7.20-7.26 (m, 2H), 7.60 (s,1H), 8.09-8.14 (m, 2H), 9.26 (b, 1H);

LC-MS (retention time: 1.91 min, method B), MS m/z 783 (M⁺+H).

Example 5 Preparation of Compound 5

Step 1:

This product was prepared by the same procedure as described in Example3, Step 1&2, except using 3-methoxy-phenethyl alcohol instead.

LC-MS (retention time: 1.03 min, method B), MS m/z 257 (M⁺+H).

Step 2:

This product was prepared by the same procedure as described in Example1, Step 2, except using the product of Example 5, Step 1 instead.

¹H NMR (CDCl₃) δ 2.72 (s, 3H), 3.90 (s, 3H), 7.00-7.02 (m, 1H), 7.41 (t,J=8.0 Hz, 1H), 7.55 (d, J=7.5 Hz, 1H), 7.59 (d, J=2.0 Hz, 1H), 7.89 (s,1H);

LC-MS (retention time: 1.89 min, method B), MS m/z 275, 277 (M⁺+H).

Step 3:

This product was prepared by the same procedure as described in Example1, Step 5, except using the product of Example 5, Step 2 instead.

¹H NMR (CD₃OD) δ 1.02 (s, 9H), 1.18 (s, 9H), 2.37-2.47 (m, 1H), 2.64 (s,3H), 2.85-2.93 (m, 1H), 3.88 (s, 3H), 4.00-4.08 (m, 1H), 4.14 (b, 1H),4.49-4.55 (m, 1H), 4.62-4.71 (m, 1H), 5.71 (b, 1H), 7.02-7.04 (m, 1H),7.40 (t, J=8.0 Hz, 1H), 7.58-7.62 (m, 3H);

LC-MS (retention time: 1.90 min, method B), MS m/z 583 (M⁺+H).

Step 4:

Compound 5 was prepared by the same procedure as described in Example 1,Step 9, except using the product of Example 5, Step 3 instead.

¹H NMR (CD₃OD) δ 1.01-1.09 (m, 12H), 1.17-1.29 (m, 10H), 1.44-1.47 (m,1H), 1.87-1.91 (m, 1H), 2.20-2.40 (m, 2H), 2.63-2.65 (m, 4H), 2.89-2.98(m, 1H), 3.89 (s, 3H), 4.08-4.20 (m, 2H), 4.44-4.65 (m, 2H), 5.13 (d,J=11.7 Hz, 1H), 5.32 (d, J=15.0 Hz, 1H), 5.72-5.85 (m, 2H), 7.02-7.05(m, 1H), 7.41 (t, J=8.0 Hz, 1H), 7.55-7.61 (m, 3H);

LC-MS (retention time: 1.96 min, method B), MS m/z 795 (M⁺+H).

Example 6 Preparation of Compound 6

Step 1:

This product was prepared by the same procedure as described in Example3, Step 1&2, except using 2-methoxy-phenethyl alcohol instead.

LC-MS (retention time: 1.10 min, method B), MS m/z 257 (M⁺+H).

Step 2:

This product was prepared by the same procedure as described in Example1, Step 2, except using the product of Example 6, Step 1 instead.

¹H NMR (CDCl₃) δ 2.721 (s, 3H), 3.88 (s, 3H), 7.03 (d, J=8.0 Hz, 1H),7.11 (t, J=7.5 Hz, 1H), 7.41-7.44 (m, 1H), 7.79-7.81 (m, 1H), 8.04 (s,1H);

LC-MS (retention time: 1.92 min, method B), MS m/z 275, 277 (M⁺+H).

Step 3:

This product was prepared by the same procedure as described in Example1, Step 5, except using the product of Example 6, Step 2 instead.

¹H NMR (CD₃OD) δ 1.02 (s, 9H), 1.20 (s, 9H), 2.37-2.47 (m, 1H), 2.63 (s,3H), 2.85-2.93 (m, 1H), 3.89 (s, 3H), 4.00-4.08 (m, 1H), 4.14 (b, 1H),4.49-4.55 (m, 1H), 4.62-4.71 (m, 1H), 5.56 (b, 1H), 7.09 (t, J=7.5 Hz,1H), 7.15 (d, J=8.5 Hz, 1H), 7.41-7.44 (m, 1H), 7.52 (s, 1H), 7.67 (d,J=8.0 Hz, 1H);

LC-MS (retention time: 1.76 min, method B), MS m/z 583 (M⁺+H).

Step 4:

Compound 6 was prepared by the same procedure as described in Example 1,Step 9, except using the product of Example 6, Step 3 instead.

¹H NMR (CD₃OD) δ 1.01-1.08 (m, 12H), 1.17-1.26 (m, 10H), 1.44-1.47 (m,1H), 1.87-1.91 (m, 1H), 2.20-2.40 (m, 2H), 2.63-2.65 (m, 4H), 2.89-2.98(m, 1H), 3.88 (s, 3H), 4.08-4.12 (m, 1H), 4.19 (b, 1H), 4.44-4.65 (m,2H), 5.13 (d, J=11.7 Hz, 1H), 5.32 (d, J=15.0 Hz, 1H), 5.59 (b, 1H),5.72-5.80 (m, 1H), 7.09 (t, J=7.5 Hz, 1H), 7.15 (d, J=8.5 Hz, 1H),7.41-7.45 (m, 1H), 7.66 (s, 1H), 7.66-7.67 (m, 1H);

LC-MS (retention time: 1.93 min, method B), MS m/z 795 (M⁺+H).

Example 7 Preparation of Compound 7

Step 1:

This product was prepared by the same procedure as described in Example1, Step 5, except using 2-chloro-quinoline instead.

LC-MS (retention time: 1.73 min, method B), MS m/z 472 (M⁺+H).

Step 2:

Compound 7 was prepared by the same procedure as described in Example 1,Step 9, except using the product of Example 7, Step 1 instead.

¹H NMR (CD₃OD) δ 1.01-1.08 (m, 12H), 1.17-1.26 (m, 10H), 1.44-1.47 (m,1H), 1.87-1.91 (m, 1H), 2.23-2.30 (m, 2H), 2.52-2.57 (m, 1H), 2.89-2.98(m, 1H), 4.10-4.14 (m, 1H), 4.09-4.15 (m, 2H), 4.47-4.51 (m, 1H), 5.13(d, J=10.0 Hz, 1H), 5.32 (d, J=17.0 Hz, 1H), 5.73-5.78 (m, 1H), 5.92 (b,1H), 6.90-6.92 (m, 1H), 7.42 (t, J=7.5 Hz, 1H), 7.64 (t, J=7.5 Hz, 1H),7.78-7.82 (m, 2H), 8.13 (d, J=7.5 Hz, 1H), 9.18 (d, 1H);

LC-MS (retention time: 1.75 min, method B), MS m/z 684 (M⁺+H).

Example 8 Preparation of Compound 8

Step 1:

This product was prepared by the same procedure as described in Example1, Step 6b through 8, without using Step 6a, an enzymatic resolutionstep.

LC-MS (retention time: 0.24 min, method B), MS m/z 231 (M⁺+H).

Step 2:

To an iced mixture of N—BOC-4-trans-hydroxy-L-proline (1.58 g, 6.83mmol), cyclopropanesulfonic acid(1-amino-2-vinyl-cyclopropanecarbonyl)-amide hydrochloride (Example 8,Step 1) (2.00 g, 7.52 mmol) and HATU (3.89 g, 10.2 mmol) in CH₂Cl₂ (100mL) was added diisopropylethylamine (4.41 g, 34.2 mmol). The formedsolution was allowed to warm up to the ambient temperature for 12 h.Diluted with EtOAc (200 mL), washed with 5% H₃PO₄ and brine, dried overMgSO₄, filtered, evaporated. The residue was purified by flashchromatography (gradient, 2:1-1:1 hexane-acetone) to yield 1.25 g (41%)of the desired product.

¹H NMR (DMSO-d₆) δ 1.00-1.08 (m, 4H), 1.34-1.40 (m, 1:2, 10H), 1.62-1.70(m, 1H), 1.76-1.87 (m, 1H), 2.02-2.21 (m, 2H), 2.81-2.95 (m, 1H),3.20-3.45 (m, 2H), 4.04-4.09 (m, 1H), 4.26 (b, 1H), 5.08-5.12 (m, 1H),5.26 (d, J=17.1 Hz, 1H), 5.59-5.69 (m, 1H), 8.59, 8.87 (rotamers, 1:2,1H), 10.48-11.15 (rotamers, 2:1, 1H);

LC-MS (retention time: 1.25 min, method B), MS m/e 444 (M⁺+H).

Step 3:

This product was prepared by the same procedure as described in Example1, Step 8, except using the product of Example 8, Step 2 instead.

LC-MS (retention time: 1.02 min, method B), MS m/e 344 (M⁺+H).

Step 4:

To an iced mixture of N—BOC-4-trans-hydroxy-L-proline (1.58 g, 6.83mmol), cyclopropanesulfonic acid(1-amino-2-vinyl-cyclopropanecarbonyl)-amide hydrochloride (Example 8,Step 3) (2.00 g, 7.52 mmol) and HATU (3.89 g, 10.2 mmol) in CH₂Cl₂ (100mL) was added diisopropylethylamine (4.41 g, 34.2 mmol). The formedsolution was allowed to warm up to the ambient temperature for 12 h.Diluted with EtOAc (200 mL), washed with 5% H₃PO₄ and brine, dried overMgSO₄, filtered, evaporated. The residue was purified by flashchromatography (gradient, 2:1-1:1 hexane-acetone) to yield 1.25 g (41%)of the desired product.

¹H NMR (CD₃OD) δ 0.99-1.07 (m, 11H), 1.35-1.44 (m, 13H), 1.75-1.87 (m,1H), 2.09-2.22 (m, 2H), 2.88-2.94 (m, 1H), 3.74-3.82 (m, 2H), 4.28-4.30(m, 1H), 4.33-4.38 (m, 1H), 4.48 (b, 1H), 5.11-5.13 (m, 1H), 5.30 (d,J=15.0 Hz, 1H), 5.70-5.78 (m, 1H), 6.51-6.61 (m, 1H);

LC-MS (retention time: 1.26 min, method B), MS m/e 557 (M⁺+H).

Step 5:

To a solution of the product of Example 8, Step 4 (56 mg, 0.1 mmol) inDMSO (2 mL) was added potassium tert-butoxide (49 mg, 0.44 mmol). Theformed solution was stirred at the ambient temperature for 1 h beforeaddition of 4-chloro-7-methyl-2-trifluoromethyl-[1,8]naphthyridine (P.Ferrarini et al, J Heterocyclic Chem, 1983, p 1053) (30 mg, 0.12 mmol).The final solution was stirred for 12 h. Quenched with iced water,acidified with 1M HCl to pH 4, extracted with EtOAc (20 mL, ×2). Theorganic layers were washed with brine, dried over MgSO₄, filtered,evaporated. The residue was purified by prep-HPLC to yield 16 mg (21%)of Compound 8 as a pink solid.

¹H NMR (CD₃OD) δ 0.92-0.99 (m, 11H), 1.01-1.04 (m, 11H), 1.22-1.45 (m,2H), 1.76-1.85 (m, 1H), 2.18-2.40 (m, 2H), 2.76 (s, 3H), 2.86-2.97 (m,1H), 4.00-4.11 (m, 2H), 4.48-4.58 (m, 2H), 5.09-5.12 (m, 1H), 5.28-5.31(m, 1H), 5.59 (b, 1H), 5.69-5.78 (m, 1H), 6.39-6.48 (m, 1H), 7.58-7.64(m, 2H), 8.08 (s, 1H), 8.64-8.68 (m, 1H), 8.85-8.91 (m, 1H);

LC-MS (retention time: 1.89 min, method B), MS m/e 767 (M⁺+H).

Example 9 Preparation of Compound 9

Compound 9 was prepared by the same procedure as described in Example 8,Step 5, except using 7-chloro-5-ethyl-3-methyl-isoxazolo[4,5-b]pyridine(R. Nesi et al, Synth Comm. 1992, 22(16), 2349) instead.

¹H NMR (CD₃OD) δ 1.01-1.09 (m, 11H), 1.21-1.25 (m, 11H), 1.36 (t, J=7.8Hz, 3H), 1.38-1.47 (m, 2H), 1.80-1.90 (m, 1H), 2.20-2.31 (m, 2H), 2.59(s, 3H), 2.90-3.00 (m, 3H), 4.01-4.18 (m, 2H), 4.41-4.51 (m, 2H),5.11-5.15 (m, 1H), 5.27-5.32 (m, 1H), 5.58 (b, 1H), 5.70-5.80 (m, 1H),7.11 (s, 1H), 7.72, 7.98 (1:1, 1H), 9.00, 9.22 (1:1, 1H);

LC-MS (retention time: 1.75 min, method B), MS m/e 717 (M⁺+H).

Example 10 Preparation of Compound 10

Compound 10 was prepared by the same procedure as described in Example8, Step 5, except using7-chloro-5-phenyl-3-methyl-isoxazolo[4,5-b]pyridine (Example 1, Step 2)instead.

¹H NMR (CD₃OD) δ 1.00-1.09 (m, 12H), 1.16-1.25 (m, 10H), 1.44-1.48 (m,1H), 1.79-1.89 (m, 1H), 2.20-2.40 (m, 2H), 2.64-2.66 (m, 4H), 2.89-2.98(m, 1H), 4.08-4.20 (m, 2H), 4.44-4.55 (m, 2H), 5.11-5.16 (m, 1H),5.27-5.31 (m, 1H), 5.72-5.74 (m, 2H), 7.20-7.35 (m, 1H), 7.46-7.51 (m,2H), 7.55-7.68 (m, 1H), 8.05-8.06 (m, 2H);

LC-MS (retention time: 1.97 min, method B), MS m/z 765 (M⁺+H).

Example 11 Preparation of Compound 11

Step 1:

To a solution of 3-methoxy cinnamic acid (11.04 g, 62 mmol) andtriethylamine (12.52 g, 124 mmol) in acetone (80 mL) was added ethylchloroformate (approximately 1.5 equivalents) dropwise at 0° C. Afterstirring at this temperature for 1 h, aqueous NaN₃ (6.40 g, 100 mmol in35 mL H₂O) was added dropwise and the reaction mixture was stirred for16 h at the ambient temperature. Water (100 mL) was added to the mixtureand the volatile was removed in vacuo. The resulting slurry wasextracted with toluene (3×50 mL) and the combined organic layers weredried over MgSO₄. This dried solution was added dropwise to a heatedsolution of diphenylmethane (50 mL) and tributylamine (30 mL) at 190° C.The toluene was distilled off as added. After complete addition, thereaction temperature was raised to 210° C. for 2 h. After cooling, theprecipitated product was collected by filtration, washed with hexane(2×50 mL), and dried to yield the desired product as a white solid (5.53g, 51%) (Nicolas Briet at el, Tetrahedron, 2002, 5761-5766).

LC-MS (retention time: 0.82 min, method B), MS m/z 176 (M⁺+H).

Step 2:

6-Methoxy-2H-isoquinolin-1-one (5.0 g, 28.4 mmol) in POCl₃ (10 mL) washeated to gentle reflux for 3 h the evaporated in vacuo (Nicolas Brietat el, Tetrahedron, 2002, 5761-5766). The residue was poured into icedwater (20 mL) and neutralized to pH 10 with 10 M NaOH. Extracted withCHCl₃. The organic layer was washed with brine, dried over MgSO₄,filtered, evaporated. The residue was purified by flash chromatography(1:1 hexane-EtOAc) to afford 4.41 g (80%) of the desired product as awhite solid.

¹H NMR (CD₃OD) δ 3.98 (s, 3H), 7.34-7.38 (m, 2H), 7.69 (d, J=5.5 Hz,1H), 8.10 (d, J=6.0 Hz, 1H), 8.23 (d, J=9.5 Hz, 1H);

LC-MS (retention time: 1.42 min, method B), MS m/z 194 (M⁺+H).

Step 3:

To a solution of N—BOC-3-(R)-hydroxy-L-proline (892 mg, 3.89 mmol) inDMSO (40 mL) at the ambient temperature was added potassiumtert-butoxide (1.34 g, 12.0 mmol) in one portion. The formed suspensionwas stirred at this temperature for 30 min before being cooled to 10° C.1-chloro-6-methoxy-isoquinoline (example 11, Step 2) (785 mg, 4.05 mmol)was added as solid in one portion and the final mixture was stirred atthe ambient temperature for 12 h. Quenched with iced 5% citric acid(aq), extracted with EtOAC (100 mL). The aqueous phase was extractedwith EtOAC again. The combined organic layers were washed with 5% citricacid (aq) and brine respectively, dried over MgSO₄, filtered. Thefiltrate was evaporated in vacuo to dryness to yield 1.49 g (99%) of thedesired product as an off-white foam. This material was used in the nextstep reaction as crude without further purification.

¹H NMR (CD₃OD) δ 1.42, 1.44 (rotamers, 9H), 2.38-2.43 (m, 1H), 2.66-2.72(m, 1H), 3.80-3.87 (m, 2H), 3.92 (s, 3H), 4.44-4.52 (m, 1H), 5.73 (b,1H), 7.16-7.18 (m, 2H), 7.24-7.25 (m, 1H), 7.87-7.88 (m, 1H), 8.07 (d,J=8.5 Hz, 1H);

LC-MS (retention time: 1.62 min, method B), MS m/z 389 (M⁺+H).

Step 4:

To a mixture of the product of Example 11, Step 3 (1.49 g, 3.84 mmol),HATU (2.19 g, 5.76 mmol), and cyclopropanesulfonic acid(1-(R)-amino-2-(S)-vinyl-cyclopropanecarbonyl)-amide HCl salt (Example1, Step 8) (1.12 g, 4.22 mmol) in CH₂Cl₂ (50 mL) was added DIPEA (1.29g, 11.5 mmol) at 0° C. After stirring at the ambient temperature for 12h, the formed solution was diluted with CH₂Cl₂ (50 mL), washed with iced5% citric acid (aq). The organic layer was washed with 5% citric acid(aq) and brine respectively, dried over MgSO₄, and filtered. Thefiltrate was evaporated in vacuo to dryness. The residue wasrecrystallized from methanol to yield 1.60 g (70%) of the desiredproduct as a white solid.

¹H NMR (CD₃OD) δ 1.05-1.08 (m, 2H), 1.16-1.20 (m, 1H), 1.24-1.27 (m,1H), 1.42-1.45 (m, 10H), 1.88 (dd, J=8.09, 5.34 Hz, 1H), 2.24-2.30 (m,2H), 2.53-2.57 (m, 1H), 2.94-2.98 (m, 1H), 3.80 (d, J=12.5 Hz, 1H),3.86-3.89 (m, 1H), 3.93 (s, 3H), 4.40-4.42 (m, 1H), 5.13 (d, J=10.5 Hz,1H), 5.32 (d, J=18.0 Hz, 1H), 5.72-5.81 (m, 2H), 7.17-7.20 (m, 2H), 7.26(d, J=6.0 Hz, 1H), 7.88 (d, J=6.0 Hz, 1H), 8.07 (d, J=9.0 Hz, 1H);

LC-MS (retention time: 1.74 min, method B), MS m/z 601 (M⁺+H).

Step 5:

To an iced solution of the product of Example 11, Step 4 (1.50 g, 2.50mmol) in CH₂Cl₂ (10 mL) was added TFA (10 mL). The formed solution wasallowed to warm to the ambient temperature for 2 h. The solvent wasremoved in vacuo. The residue was triturated with 1M HCl in ether.Filtered, washed with ether to yield 1.43 g (99.8%) of the desiredproduct as a hygroscopic white solid.

¹H NMR (CD₃OD) δ 1.03-1.208 (m, 4H), 1.26-1.31 (m, 1H), 1.37-1.40 (m,1H), 1.95-1.97 (m, 1H), 2.32-2.37 (m, 1H), 2.42-2.48 (m, 1H), 2.95-2.99(m, 1H), 3.88 (d, J=12.5 Hz, 2H), 3.98 (s, 3H), 4.40-4.42 (m, 1H), 5.16(d, J=10.5 Hz, 1H), 5.33 (d, J=18.0 Hz, 1H), 5.62-5.69 (m, 1H), 5.97 (b,1H), 7.30-7.34 (m, 2H), 7.47 (d, J=6.0 Hz, 1H), 7.90 (d, J=6.5 Hz, 1H),8.34 (d, J=9.0 Hz, 1H), 9.14 (b, 1H);

LC-MS (retention time: 1.12 min, method B), MS m/z 501 (M⁺+H).

Step 6:

To a mixture of the product of Example 11, Step 5 (1.49 g, 3.84 mmol),HATU (2.19 g, 5.76 mmol), and N—BOC-t-butyl-L-glycine (1.12 g, 4.22mmol) in CH₂Cl₂ (50 mL) was added DIPEA (1.29 g, 11.5 mmol) at 0° C.After stirring at the ambient temperature for 12 h, the formed solutionwas diluted with CH₂Cl₂ (50 mL), washed with iced 5% citric acid (aq).The organic layer was washed with 5% citric acid (aq) and brinerespectively, dried over MgSO₄, and filtered. The filtrate wasevaporated in vacuo to dryness. The residue was purified by prep-HPLC(40% B to 100% B, 15 min gradient time) to yield 1.60 g (70%) ofCompound 11 as a white solid.

¹H NMR (CD₃OD) δ 1.00-1.08 (m, 12H), 1.23-1.25 (m, 1H), 1.27 (s, 9H),1.40-1.45 (m, 1H), 1.85-1.88 (m, 1H), 2.20-2.30 (m, 2H), 2.55-2.61 (m,1H), 2.91-2.97 (m, 1H), 3.92 (s, 3H), 4.02-4.06 (m, 1H), 4.21-4.24 (m,1H), 4.40-4.42 (m, 1H), 4.49-4.51 (m, 1H), 5.12 (d, J=10.5 Hz, 1H), 5.28(d, J=18.0 Hz, 1H), 5.69-5.74 (m, 1H), 5.81 (b, 1H), 6.60 (d, J=10.0 Hz,1H), 7.08-7.10 (m, 1H), 7.18 (s, 1H), 7.25 (d, J=6.0 Hz, 1H), 7.88 (d,J=6.0 Hz, 1H), 8.09 (d, J=9.0 Hz, 1H);

LC-MS (retention time: 1.75 min, method B), MS m/z 714 (M⁺+H);

Anal. Calcd for C₃₅H₄₇N₅O₉S.0.5 H₂O: C, 58.16; H, 6.69; N, 9.69. Found:C, 58.01; H, 6.46; N, 9.55.

Step 7:

To a solution of Compound 11 (71 mg, 0.1 mmol) in CH₂Cl₂ (5 mL) at −78°C. was added 1M HCl in ether (0.2 mL, 0.2 mmol). After stirring at thistemperature for 10 min, the volatile was removed in vacuo withoutheating bath. The residue was triturated with ether, filtered, washedwith ether and dried to yield 61 mg (85%) of the desired HCl salt ofCompound 11 as a very fine solid.

¹H NMR (CD₃OD) δ 1.00-1.08 (m, 12H), 1.19 (s, 9H), 1.23-1.25 (m, 1H),1.40-1.45 (m, 1H), 1.85-1.91 (m, 1H), 2.20-2.26 (m, 1H), 2.31-2.42 (m,1H), 2.65-2.78 (m, 1H), 2.92-2.97 (m, 1H), 4.00 (s, 3H), 4.10-4.16 (m,2H), 4.51-4.64 (m, 2H), 5.13 (d, J=10.5 Hz, 1H), 5.30 (d, J=18 Hz, 1H),5.69-5.79 (m, 1H), 5.84 (b, 1H), 7.28 (d, J=9.3 Hz, 1H), 7.40 (s, 1H),7.55 (d, J=6.3 Hz, 1H), 7.89-7.92 (m, 1H), 8.29 (d, J=9.0 Hz, 1H), 9.21(b, 1H);

LC-MS (retention time: 1.75 min, method B), MS m/z 714 (M⁺+H).

Anal. Calcd for C₃₅H₄₇N₅O₉S.1.0HCl: C, 56.02; H, 6.44; N, 9.33; Cl,4.72: S, 4.27.

Found: C, 55.80; H, 6.42; N, 9.15; Cl, 4.56: S, 4.09.

Step 8:

To a 25 ml 2 neck flask was added a stir bar, septa and N₂ gas adapter.Compound 11 (99.7 mg, 0.140 mmol) was weighed out and added to thereaction flask. The reaction flask was purged and placed under a N₂atmosphere. 850 ul of acetone was added to the flask to provide a clearsolution. To this solution at room temperature was added 780 ul of a0.179 M solution of KOH (aq.) prepared by the dissolution of solid KOH(502.8 mg, 8.97 mmol) in 50 ml of H₂O. The solution warmed slightly uponaddition of the KOH but remained clear. The clear solution was allowedto stir at RT for 2 hours. The product crystallized out of solution andwas isolated by filtration. The cake was washed with cold acetone toafford 42 mg (40% yield) of the desired product as fine white needles:¹H NMR (DMSO) δ 0.68 (m, 1H), 0.72 (m, 1H), 0.88 (s, 1H), 0.92 (s, 1H),1.24 (s, 1H), 1.38 (s, 1H), 1.50 (b, 1H), 1.81 (b, 1H), 2.68 (b, 2H),3.90 (s, 3H), 3.95-4.10 (m, 3H), 4.40 (t, J=10 Hz, 1H), 4.85 (m, 1H),5.04 (m, 1H), 5.71 (b, 1H), 6.01 (b, 1H), 6.64 (d, J=10 Hz, 1H), 7.10(m, 1H), 7.30 (m, J=5 Hz, 2H), 7.95 (d, J=10 Hz, 1H), 8.08 (d, J=15 Hz,1H).

Elemental analysis for C₃₅H₄₆KN₅O₉S.H₂O; calc. C, 54.60; H, 6.28; K,5.08; N, 9.10 actual C, 54.88; H, 6.23; K, 5.05; N, 9.01; MS m/e 714(MH⁺);

Example 12 Preparation of Compound 12

Compound 12 was prepared by the same procedure as described in Example11, Step 6, except using N—BOC-L-valine instead.

¹H NMR (CD₃OD) δ 0.94-0.98 (m, 6H), 1.07-1.09 (m, 3H), 1.21-1.25 (m,10H), 1.40-1.43 (m, 1H), 1.88-1.89 (m, 1H), 2.05-2.09 (m, 1H), 2.22-2.35(m, 2H), 2.57-2.61 (m, 1H), 2.94-2.97 (m, 1H), 3.92 (s, 3H), 4.03-4.06(m, 2H), 4.47-4.55 (m, 2H), 5.12 (d, J=10.5 Hz, 1H), 5.32 (d, J=18.1 Hz,1H), 5.74-5.81 (m, 1H), 5.86 (b, 1H), 7.10 (d, J=9.0 Hz, 1H), 7.18 (s,1H), 7.25 (d, J=6.0 Hz, 1H), 7.88 (d, J=6.0 Hz, 1H), 8.10 (d, J=9.0 Hz,1H);

LC-MS (retention time: 1.71 min, method B), MS m/z 700 (M⁺+H).

Example 13 Preparation of Compound 13

Compound 13 was prepared by the same procedure as described in Example11, Step 6, except using N—BOC-L-alloisoleucine instead.

¹H NMR (CD₃OD) δ 0.89-0.96 (m, 6H), 1.07-1.18 (m, 5H), 1.28 (s, 9H),1.42-1.45 (m, 1H), 1.50-1.54 (m, 1H), 1.87-1.89 (m, 2H), 2.23-2.34 (m,2H), 2.57-2.61 (m, 1H), 2.92-2.95 (m, 1H), 3.92 (s, 3H), 4.05-4.07 (m,1H), 4.22-4.24 (m, 1H), 4.37-4.40 (m, 1H), 4.54-4.56 (m, 1H), 5.13 (d,J=10.5 Hz, 1H), 5.32 (d, J=18.0 Hz, 1H), 5.75-5.82 (m, 1H), 5.86 (b,1H), 7.12 (d, J=9.0 Hz, 1H), 7.19 (s, 1H), 7.24 (d, J=6.0 Hz, 1H), 7.88(d, J=6.0 Hz, 1H), 8.10 (d, J=9.0 Hz, 1H);

LC-MS (retention time: 1.77 min, method B), MS m/z 714 (M⁺+H).

Example 14 Preparation of Compound 14

Step 1:

A mixture of Compound 11 (150 mg, 0.21 mmol) and Pearlmann's catalyst(Pd(OH)₂, 15 mg) in EtOAc (10 mL) was placed on Parr shaker for 20 minunder 10 psi H₂. Filtered through celite. The filtrate was evaporated invacuo. The residue was purified by prep-HPLC to provide 67 mg (45%) ofCompound 14 as a white solid.

¹H NMR (CD₃OD) δ 0.96-0.99 (m, 4H), 1.04 (s, 9H), 1.07-1.09 (m, 2H),1.21-1.24 (m, 2H), 1.27 (s, 9H), 1.51-1.65 (m, 4H), 2.25-2.27 (m, 1H),2.55-2.61 (m, 1H), 2.94-2.98 (m, 1H), 3.92 (s, 3H), 4.02-4.06 (m, 1H),4.21-4.24 (m, 1H), 4.40-4.42 (m, 1H), 4.49-4.51 (m, 1H), 5.81 (b, 1H),6.59 (d, J=10.0 Hz, 1H), 7.08-7.10 (m, 1H), 7.18 (d, J=1.5 Hz, 1H), 7.24(d, J=6.0 Hz, 1H), 7.88 (d, J=6.0 Hz, 1H), 8.08 (d, J=9.0 Hz, 1H);

LC-MS (retention time: 1.76 min, method B), MS m/z 716 (M⁺+H).

Example 15 Preparation of Compound 15

Compound 15 was isolated form the same reaction of making Compound 14with a slightly longer retention time as a by-product in 15% yield.

¹H NMR (CD₃OD) δ 0.92-1.10 (m, 17H), 1.26-1.36 (m, 13H), 1.64-1.72 (m,1H), 1.90-1.96 (m, 1H), 2.30-2.40 (m, 1H), 2.63-2.67 (m, 1H), 2.96-3.00(m, 1H), 3.92 (s, 3H), 4.03-4.07 (m, 1H), 4.24 (b, 1H), 4.40-4.42 (m,1H), 4.49-4.51 (m, 1H), 5.83 (b, 1H), 7.08-7.11 (m, 1H), 7.19 (d, J=2.0Hz, 1H), 7.25 (d, J=6.0 Hz, 1H), 7.89 (d, J=6.0 Hz, 1H), 8.10 (d, J=9.0Hz, 1H), 8.51 (b, 1H);

LC-MS (retention time: 1.83 min, method B), MS m/z 718 (M⁺+H).

Example 16 Preparation of Compound 16

Step 1:

To a solution of Compound 11 (420 mg, 0.59 mmol) in DCM (5 mL) at 0° C.was added TFA (5 mL). After stirring at this temperature for 2 h, thevolatile was removed in vacuo. The residue was triturated with 1M HCl inether (5 mL), filtered, washed with ether and dried to yield 360 mg(89%) of the desired HCl salt as a very fine solid.

LC-MS (retention time: 1.28 min, method B), MS m/z 614 (M⁺+H).

Step 2:

To a suspension of the product of Example 16, Step 1 (39 mg, 0.06 mmol),and DIPEA (20 mg, 0.18 mmol) in DCM (1 mL) at 0° C. was added methylchloroformate (6.8 mg, 0.072 mmol). After stirring at this temperaturefor 2 h, the volatile was removed in vacuo. The residue was purified byprep-HPLC to give 21 mg (58%) of Compound 16 as a white crystal.

¹H NMR (CD₃OD) δ 1.05-1.09 (m, 11H), 1.22-1.25 (m, 2H), 1.41-1.44 (m,1H), 1.86-1.89 (m, 1H), 2.22-2.32 (m, 2H), 2.59-2.63 (m, 1H), 2.89-2.93(m, 1H), 3.48 (s, 3H), 3.92 (s, 3H), 4.06-4.10 (m, 1H), 4.31-4.33 (m,1H), 4.38-4.40 (m, 1H), 4.50-4.52 (m, 1H), 5.12 (d, J=10.5 Hz, 1H), 5.30(d, J=18.0 Hz, 1H), 5.71-5.80 (m, 1H), 5.85 (b, 1H), 6.95 (d, J=10.0 Hz,1H), 7.13-7.16 (m, 1H), 7.19 (s, 1H), 7.25 (d, J=6.0 Hz, 1H), 7.88 (d,J=6.0 Hz, 1H), 8.09 (d, J=9.0 Hz, 1H);

LC-MS (retention time: 1.54 min, method B), MS m/z 672 (M⁺+H).

Example 17 Preparation of Compound 17

Compound 17 was prepared by the same procedure as described in Example16, Step 2, except using isopropyl chloroformate instead.

¹H NMR (CD₃OD) δ 1.00-1.09 (m, 15H), 1.13-1.16 (m, 2H), 1.24-1.26 (m,2H), 1.40-1.45 (m, 1H), 1.86-1.89 (m, 1H), 2.21-2.31 (m, 2H), 2.55-2.61(m, 1H), 2.91-2.97 (m, 1H), 3.92 (s, 3H), 4.04-4.08 (m, 1H), 4.30 (b,1H), 4.40 (d, J=10 Hz, 1H), 4.49-4.54 (m, 2H), 5.12 (d, J=10.5 Hz, 1H),5.29 (d, J=18.0 Hz, 1H), 5.71-5.77 (m, 1H), 5.84 (b, 1H), 6.80 (d,J=10.0 Hz, 1H), 7.11 (d, J=9.0 Hz, 1H), 7.19 (s, 1H), 7.25 (d, J=6.0 Hz,1H), 7.88 (d, J=6.0 Hz, 1H), 8.08 (d, J=9.0 Hz, 1H);

LC-MS (retention time: 1.74 min, method B), MS m/z 700 (M⁺+H).

Example 18 Preparation of Compound 18

Compound 18 was prepared by the same procedure as described in Example16, Step 2, except using neopentyl chloroformate instead.

¹H NMR (CD₃OD) δ 0.61 (b, 1H), 0.84 (s, 8H), 1.05-1.09 (m, 11H),1.23-1.25 (m, 2H), 1.39-1.44 (m, 1H), 1.85-1.88 (m, 1H), 2.20-2.30 (m,2H), 2.56-2.62 (m, 1H), 2.91-2.97 (m, 1H), 3.38 (d, J=9.0 Hz, 1H), 3.55(d, J=9.0 Hz, 1H), 3.92 (s, 3H), 4.02-4.06 (m, 1H), 4.32 (d, J=9.5 Hz,1H), 4.41 (d, J=9.0 Hz, 1H), 4.49-4.51 (m, 1H), 5.12 (d, J=10.5 Hz, 1H),5.28 (d, J=18.0 Hz, 1H), 5.69-5.74 (m, 1H), 5.81 (b, 1H), 6.90 (d,J=10.0 Hz, 1H), 7.08-7.10 (m, 1H), 7.19 (s, 1H), 7.26 (d, J=6.0 Hz, 1H),7.88 (d, J=6.0 Hz, 1H), 8.07 (d, J=9.0 Hz, 1H);

LC-MS (retention time: 1.84 min, method B), MS m/z 728 (M⁺+H).

Example 19 Preparation of Compound 19

Compound 19 was prepared by the same procedure as described in Example16, Step 2, except using (S)-3-furanochloroformate (J. Campbell, A.Good, WO 20020808) instead.

¹H NMR (CD₃OD) δ 1.03-1.08 (m, 11H), 1.23-1.26 (m, 2H), 1.38-1.46 (m,1H), 1.64-1.71 (m, 1H), 1.85-1.90 (m, 2H), 2.20-2.30 (m, 2H), 2.55-2.61(m, 1H), 2.91-2.97 (m, 1H), 3.66-3.72 (m, 4H), 3.93 (s, 3H), 4.05-4.09(m, 1H), 4.27-4.29 (m, 1H), 4.40-4.42 (m, 1H), 4.55-4.59 (m, 1H),4.75-4.77 (m, 1H), 5.12 (d, J=10.5 Hz, 1H), 5.28 (d, J=18 Hz, 1H),5.73-5.80 (m, 1H), 5.85 (b, 1H), 7.06 (d, J=10.0 Hz, 1H), 7.13 (d, J=9.0Hz, 1H), 7.20 (s, 1H), 7.25 (d, J=6.0 Hz, 1H), 7.89 (d, J=6.0 Hz, 1H),8.07 (d, J=9.0 Hz, 1H);

LC-MS (retention time: 1.52 min, method B), MS m/z 728 (M⁺+H).

Example 20 Preparation of Compound 20

Step 1:

This product was prepared by the same procedure as described in Example11, Step 2, except using 6-chloro-2H-isoquinolin-1-one ((Nicolas Brietat el, Tetrahedron, 2002, 5761-5766) instead.

LC-MS (retention time: 1.07 min, method B), MS m/z 180 (M⁺+H).

Step 2:

This product was prepared by the same procedure as described in Example1, Step 5, except using the product of Example 20, Step 1 instead.

¹H NMR (CD₃OD) δ 1.04 (s, 9H), 1.20 (s, 9H), 2.36-2.41 (m, 1H),2.74-2.78 (m, 1H), 4.01-4.04 (m, 1H), 4.19-4.21 (m, 1H), 4.47-4.49 (m,1H), 4.67-4.70 (m, 1H), 5.84 (b, 1H), 7.28 (d, J=6.0 Hz, 1H), 7.47 (d,J=6.0 Hz, 1H), 7.84 (s, 1H), 8.00 (d, J=6.0 Hz, 1H), 8.20 (d, J=9.0 Hz,1H);

LC-MS (retention time: 1.88 min, method B), MS m/z 506 (M⁺+H).

Step 3:

Compound 20 was prepared by the same procedure as described in Example1, Step 9, except using the product of Example 20, Step 2 instead.

¹H NMR (CD₃OD) δ 0.99-1.11 (m, 12H), 1.20-1.26 (m, 10H), 1.43-1.46 (m,1H), 1.87-1.90 (m, 1H), 2.22-2.31 (m, 2H), 2.60-2.64 (m, 1H), 2.92-2.97(m, 1H), 4.06-4.08 (m, 1H), 4.21-4.23 (m, 1H), 4.45-4.47 (m, 1H),4.53-4.56 (m, 1H), 5.13 (d, J=10.5 Hz, 1H), 5.29 (d, J=18.0 Hz, 1H),5.72-5.80 (m, 1H), 5.88 (b, 1H), 6.58 (d, J=10.0 Hz, 1H), 7.29 (d, J=6.0Hz, 1H), 7.47 (d, J=9.0 Hz, 1H), 7.86 (s, 1H), 8.01 (d, J=6.0 Hz, 1H),8.18 (d, J=9.0 Hz, 1H);

LC-MS (retention time: 1.94 min, method B), MS m/z 718 (M⁺+H).

Example 21 Preparation of Compound 21

Step 1:

To a mixture of the product of Example 1, Step 4 (3.00 g, 8.72 mmol),HATU (4.97 g, 13.1 mmol), and product of Example 1, Step 8 (2.55 g, 9.59mmol) in CH₂Cl₂ (100 mL) was added DIPEA (3.02 g, 27.0 mmol) at 0° C.After stirring at the ambient temperature for 12 h, the formed solutionwas diluted with CH₂Cl₂ (100 mL), washed with iced 5% citric acid (aq).The organic layer was washed with 5% citric acid (aq) and brinerespectively, dried over MgSO₄, and filtered. The filtrate wasevaporated in vacuo to dryness. The residue was purified by flash column(1:1 hexane:acetone) to yield 3.64 g (75%) of the desired product as afoam.

LC-MS (retention time: 1.41 min, method B), MS m/z 557 (M⁺+H).

Step 2:

To an iced solution of 6-bromoisoquinoline (4.16 g, 20 mmol) in CH₂Cl₂(100 mL) was added mCPBA (9.38 g, 77% pure, 42 mmol) as solid in oneportion. After stirring at the ambient temperature for 12 h, dilutedwith CH₂Cl₂ (100 mL) and washed with 1M NaOH (100 mL, ×2) and brine. Theorganic layer was dried over MgSO₄, filtered, evaporated to dryness toyield 3.83 g (86%) of the desired product as a white solid. Thismaterial was used as crude without further purification.

LC-MS (retention time: 0.77 min, method B), MS m/z 224, 226 (M⁺+H).

Step 3:

A mixture of 6-bromo-isoquinoline 2-oxide (88 mg, 0.2 mmol), pyrazole(68 mg, 1.0 mmol), CuBr (57 mg, 0.4 mmol) and cesium carbonate (130 mg,0.4 mmol) in DMF (2 mL) was heated to 140° C. for 4 h in a sealed tube.After filtration, the filtrated was purified by prep-HPLC to yield 41 mg(98%) of the desired product as an off-white solid.

¹H NMR (CDCl₃) δ 6.58-6.59 (m, 1H), 7.82 (d, J=1.0 Hz, 1H), 7.89 (d,J=7.0 Hz, 1H), 8.02 (d, J=9.0 Hz, 1H), 8.11 (d, J=2.5 Hz, 1H), 8.18-8.22(m, 2H), 8.29 (d, J=7.0 Hz, 1H), 9.07 (b, 1H);

LC-MS (retention time: 0.77 min, method B), MS m/z 212 (M⁺+H).

Step 4:

This product was prepared by the same procedure as described in Example11, Step 2 as an off-white solid, except using 6-pyrazol-isoquinoline2-oxide instead.

¹H NMR (CD₃OD) δ 7.82-7.83 (m, 2H), 8.23-8.32 (m, 4H), 8.44-8.49 (m,2H);

LC-MS (retention time: 1.35 min, method B), MS m/z 230, (M⁺+H).

Step 5:

To a solution of product of Example 21, Step 1 (45 mg, 0.08 mmol) inDMSO (2 mL) was added potassium tert-butoxide (41 mg, 0.37 mmol). Theformed solution was stirred at the ambient temperature for 30 min beforeaddition of 1-chloro-6-pyrazol-1-yl-isoquinoline (17 mg, 0.07 mmol). Thefinal solution was stirred for 12 h. Quenched with iced water, acidifiedwith 1M HCl to pH 4, extracted with EtOAc (20 mL, ×2). The organiclayers were washed with brine, dried over MgSO₄, filtered, evaporated.The residue was purified by prep-HPLC to yield 10 mg (16%) of Compound21 as a pink solid.

¹H NMR (CD₃OD) δ 1.04-1.10 (m, 12H), 1.23-1.27 (m, 10H), 1.43-1.47 (m,1H), 1.87-1.91 (m, 1H), 2.22-2.29 (m, 2H), 2.61-2.68 (m, 1H), 2.92-2.98(m, 1H), 4.07-4.11 (m, 1H), 4.24 (b, 1H), 4.46-4.60 (m, 2H), 5.13 (d,J=10.5 Hz, 1H), 5.29 (d, J=18 Hz, 1H), 5.70-5.83 (m, 1H), 5.89 (b, 1H),6.59-6.61 (m, 1H), 7.40 (d, J=10.0 Hz, 1H), 7.80 (d, J=2.5 Hz, 1H), 8.01(d, J=10.0 Hz, 2H), 8.15 (s, 1H), 8.31 (d, J=15.0 Hz, 1H), 8.42 (d,J=4.5 Hz, 1H);

LC-MS (retention time: 1.77 min, method B), MS m/z 750 (M⁺+H).

Example 22 Preparation of Compound 22

Step 1:

This product was prepared by the same procedure as described in Example11, Step 2 as an off-white solid, except using 6-bromo-isoquinoline2-oxide instead.

¹H NMR (CD₃OD) δ 7.73 (d, J=5.5 Hz, 1H), 7.85-7.91 (m, 1H), 8.22-8.31(m, 3H);

LC-MS (retention time: 1.53 min, method B), MS m/z 241, 243, 245 (M⁺+H).

Step 2:

Compound 22 was prepared by the same procedure as described in Example21, Step as a white solid, except using 1-chloro-6-bromo-isoquinolineinstead.

¹H NMR (CD₃OD) δ 0.99-1.09 (m, 12H), 1.22-1.27 (m, 10H), 1.40-1.47 (m,1H), 1.86-1.91 (m, 1H), 2.20-2.34 (m, 2H), 2.57-2.66 (m, 1H), 2.90-2.97(m, 1H), 4.05-4.09 (m, 1H), 4.21 (b 1H), 4.44-4.57 (m, 2H), 5.13 (d,J=10.5 Hz, 1H), 5.29 (d, J=18.0 Hz, 1H), 5.70-5.82 (m, 1H), 5.88 (b,1H), 7.29 (d, J=9.5 Hz, 1H), 7.60-7.63 (m, 1H), 8.00-8.12 (m, 3H);

LC-MS (retention time: 1.90 min, method B), MS m/z 762, 764 (M⁺+H).

Example 23 Preparation of Compound 23

Step 1:

This product was prepared by the same procedure as described in Example11, Step 3 as a white solid, except using 1-chloro-isoquinoline instead.

¹H NMR (CD₃OD) δ 1.42, 1.44 (rotamers, 9H), 2.39-2.44 (m, 1H), 2.68-2.72(m, 1H), 3.80-3.87 (m, 2H), 4.44-4.52 (m, 1H), 5.78 (b, 1H), 7.32-7.33(m, 1H), 7.58 (t, J=7.8 Hz, 1H)), 7.71 (t, J=7.5 Hz, 1H), 7.81 (d, J=8.0Hz, 1H), 7.95 (d, J=6.0 Hz, 1H), 8.19 (d, J=8.0 Hz, 1H);

LC-MS (retention time: 1.61 min, method B), MS m/z 359 (M⁺+H).

Step 2:

This product was prepared by the same procedure as described in Example11, Step 4, except using the product of Example 23, Step 1 instead.

¹H NMR (DMSO-d6) δ 1.00-1.09 (m, 4H), 1.35-1.38 (m, 10H), 1.69-1.84 (m,1H), 2.11-2.66 (m, 3H), 2.89-2.93 (m, 1H), 3.62-3.89 (m, 2H), 4.31 (t,J=8.1 Hz, 1H), 5.12 (d, J=10.8 Hz, 1H), 5.27 (d, J=16.8 Hz, 1H),5.58-5.70 (m, 1H), 5.76 (b, 1H), 7.43 (d, J=5.7 Hz, 1H), 7.66 (t, J=7.4Hz, 1H), 7.79 (t, J=7.5 Hz, 1H), 7.92 (d, J=8.1 Hz, 1H), 8.02 (d, J=10.0Hz, 1H), 8.13 (d, J=8.1 Hz, 1H), 9.02 (b, 1H);

LC-MS (retention time: 1.72 min, method B), MS m/z 571 (M⁺+H).

Step 3:

This product was prepared by the same procedure as described in Example11, Step 5, except using the product of Example 23, Step 2 instead.

LC-MS (retention time: 1.16 min, method B), MS m/z 471 (M⁺+H).

Step 4:

Compound 23 was prepared by the same procedure as described in Example11, Step 6 as a white solid, except using the product of Example 23,Step 3 instead.

¹H NMR (CD₃OD) δ 1.00-1.09 (m, 12H), 1.25-1.27 (m, 10H), 1.42-1.46 (m,1H), 1.86-1.90 (m, 1H), 2.22-2.34 (m, 2H), 2.60-2.67 (m, 1H), 2.92-2.99(m, 1H), 4.06-4.11 (m, 1H), 4.26 (b, 1H), 4.45-4.57 (m, 2H), 5.12 (d,J=10.2 Hz, 1H), 5.27 (d, J=16.8 Hz, 1H), 5.70-5.82 (m, 1H), 5.88 (b,1H), 7.32 (d, J=6.0 Hz, 1H), 7.52 (t, J=7.4 Hz, 1H), 7.70 (t, J=7.5 Hz,1H), 7.80 (d, J=8.1 Hz, 1H), 7.97 (d, J=6 Hz, 1H), 8.20 (d, J=8.4 Hz,1H), 9.18 (b, 1H);

LC-MS (retention time: 1.80 min, method B), MS m/z 684 (M⁺+H).

Example 24 Preparation of Compound 24

Step 1:

To a solution of N—BOC-3-(R)-hydroxy-L-proline (6.22 g, 26.9 mmol) inDMF (250 mL) at 0° C. was added NaH (60%, 3.23 g, 80.8 mmol) by severalportions. The formed suspension was stirred at this temperature for 30min. 1,3-dichloro-isoquinoline (5.33 g, 26.9 mmol) was added as solid inone portion and the final mixture was stirred at the ambient temperaturefor 12 h. Quenched with iced 5% citric acid (aq), extracted with EtOAC(300 mL). The aqueous phase was extracted with EtOAC again. The combinedorganic layers were washed with 5% citric acid (aq) and brinerespectively, dried over MgSO₄, filtered. The filtrate was evaporated invacuo to dryness to yield 10.53 g (99.8%) of4-(6-methoxy-isoquinolin-1-yloxy)-pyrrolidine-1,2-dicarboxylic acid1-tert-butyl ester as an off-white foam. This material was used in thenext step reaction as crude without further purification.

¹H NMR (CD₃OD) δ 1.43, 1.44 (rotamers, 9H), 2.39-2.44 (m, 1H), 2.68-2.72(m, 1H), 3.80-3.90 (m, 2H), 4.44-4.52 (m, 1H), 5.77 (b, 1H), 7.39 (s,1H), 7.58 (t, J=7.3 Hz, 1H), 7.71-7.78 (m, 2H), 8.16 (d, J=7.5 Hz, 1H);

LC-MS (retention time: 1.80 min, method B), MS m/z 392 (M⁺+H).

Step 2:

This product was prepared by the same procedure as described in Example11, Step 4, except using the product of Example 24, Step 1 instead.

¹H NMR (CD₃OD) δ 1.02-1.08 (m, 2H), 1.18-1.26 (m, 2H), 1.44-1.48 (m,10H), 1.84-1.91 (m, 1H), 2.22-2.36 (m, 2H), 2.57-2.60 (m, 1H), 2.95-2.99(m, 1H), 3.81-3.93 (m, 2H), 4.38-4.41 (m, 1H), 5.13 (d, J=10.8 Hz, 1H),5.31 (d, J=16.8 Hz, 1H), 5.75-5.82 (m, 2H), 7.41 (s, 1H), 7.59 (t, J=7.4Hz, 1H), 7.74-7.79 (m, 2H), 8.16 (d, J=8.0 Hz, 1H);

LC-MS (retention time: 1.82 min, method B), MS m/z 605 (M⁺+H).

Step 3:

This product was prepared by the same procedure as described in Example11, Step 5, except using the product of Example 24, Step 2 instead.

LC-MS (retention time: 1.30 min, method B), MS m/z 505 (M⁺+H).

Step 4:

Compound 24 was prepared by the same procedure as described in Example11, Step 6 as a white solid, except using the product of Example 24,Step 3 instead.

¹H NMR (CD₃OD) δ 0.99-1.09 (m, 12H), 1.22-1.29 (m, 10H), 1.42-1.46 (m,1H), 1.86-1.90 (m, 1H), 2.21-2.34 (m, 2H), 2.62-2.66 (m, 1H), 2.92-2.99(m, 1H), 4.06-4.11 (m, 1H), 4.26 (b, 1H), 4.46-4.56 (m, 2H), 5.13 (d,J=10.5 Hz, 1H), 5.29 (d, J=17.2 Hz, 1H), 5.72-5.79 (m, 1H), 5.89 (b,1H), 7.40 (d, J=6.0 Hz, 1H), 7.52 (t, J=7.4 Hz, 1H), 7.72-7.76 (m, 2H),8.18 (d, J=8.5 Hz, 1H);

LC-MS (retention time: 1.95 min, method B), MS m/z 718 (M⁺+H).

Example 25 Compound 25

Step 1:

A mixture of Example 24, Step 1 (39 mg, 0.10 mmol), phenylboronic acid(14.6 mg, 0.12 mmol), sodium tert-butoxide (38 mg, 0.40 mmol) and((t-Bu)₂POH)₂PdCl₂ (POPd) (5 mg, 0.01 mmol) in THF (2 mL) was heated toreflux for 4 h. After cooling down, the formed mixture was quenched with5% citric acid (aq) and extracted with EtOAc (20 mL). The organic layerwas washed with brine, dried over MgSO₄, filtered, evaporated. Theresidue was purified by prep-HPLC to yield 36 mg (83%) of the desiredproduct as an off-white foam.

¹H NMR (CD₃OD) δ 1.43, 1.45 (rotamers, 9H), 2.51-2.56 (m, 1H), 2.74-2.82(m, 1H), 3.88-3.92 (m, 1H), 3.98-4.01 (m, 1H), 4.50-4.57 (m, 1H), 5.95(b, 1H), 7.36-7.39 (m, 1H), 7.45-7.48 (m, 2H), 7.55 (t, J=7.3 Hz, 1H),7.70 (t, J=7.5 Hz, 1H), 7.84-7.89 (m, 2H), 8.14-8.17 (m, 3H), 9.05 (b,1H);

LC-MS (retention time: 1.97 min, method B), MS m/z 435 (M+H).

Step 2:

This product was prepared by the same procedure as described in Example11, Step 4, except using the product of Example 25, Step 1 instead.

¹H NMR (DMSO-d6) δ 0.98-1.10 (m, 4H), 1.38-1.41 (m, 10H), 1.74-1.81 (m,1H), 2.18-2.34 (m, 2H), 2.47-2.49 (m, 1H), 2.95-2.99 (m, 1H), 3.74-3.96(m, 2H), 4.34-4.37 (m, 1H), 5.12 (d, J=10.5 Hz, 1H), 5.26 (d, J=17.8 Hz,1H), 5.75-5.82 (m, 1H), 5.95 (b, 1H), 7.41-7.45 (m, 1H), 7.51-7.54 (m,2H), 7.61-7.64 (m, 1H), 7.78-7.82 (m, 1H), 7.98 (d, J=9.0 Hz, 1H), 8.06(s, 1H), 8.13-8.14 (m, 1H), 8.18-8.20 (m, 2H), 9.05 (b, 1H), 10.34 (b,1H);

LC-MS (retention time: 1.99 min, method B), MS m/z 647 (M⁺+H).

Step 3:

This product, was prepared by the same procedure as described in Example11, Step 5 as a white solid, except using the product of Example 25,Step 2 instead.

LC-MS (retention time: 1.55 min, method B), MS m/z 547 (M⁺+H).

Step 4:

Compound 25 was prepared by the same procedure as described in Example11, Step 6 as a white solid, except using the product of Example 25,Step 3 instead.

¹H NMR (CD₃OD) δ 0.92-1.09 (m, 12H), 1.26-1.30 (m, 10H), 1.43-1.46 (m,1H), 1.87-1.90 (m, 1H), 2.21-2.26 (m, 1H), 2.36-2.41 (m, 1H), 2.70-2.75(m, 1H), 2.93-2.97 (m, 1H), 4.18-4.30 (m, 2H), 4.46-4.48 (m, 1H),4.55-4.58 (m, 1H), 5.12 (d, J=10.5 Hz, 1H), 5.29 (d, J=18.0 Hz, 1H),5.72-5.79 (m, 1H), 6.10 (b, 1H), 7.37-7.40 (m, 1H), 7.46-7.49 (m, 3H),7.70 (t, J=7.5 Hz, 1H), 7.85-7.89 (m, 2H), 8.16-8.20 (m, 3H);

LC-MS (retention time: 2.08 min, method B), MS m/z 760 (M⁺+H).

Example 26 Preparation of Compound 26

Step 1:

This product was prepared by the same procedure as described in Example25, Step 1, except using 4-methoxyphenylboronic acid instead.

¹H NMR (CD₃OD) δ 1.40, 1.45 (rotamers, 9H), 2.50-2.55 (m, 1H), 2.73-2.81(m, 1H), 3.81-3.89 (m, 4H), 3.98-4.01 (m, 1H), 4.50-4.57 (m, 1H), 5.93(b, 1H), 7.02 (d, J=9.0 Hz, 2H), 7.50 (t, J=7.3 Hz, 1H), 7.67 (t, J=7.5Hz, 1H), 7.73 (s, 1H), 7.83 (d, J=8.5 Hz, 1H), 8.09 (d, J=8.5 Hz, 2H),8.15 (d, J=8.0 Hz, 1H);

LC-MS (retention time: 2.00 min, method B), MS m/z 465 (M⁺+H).

Step 2:

This product was prepared by the same procedure as described in Example11, Step 4, except using the product of Example 26, Step 1 instead.

¹H NMR (CD₃OD) δ 1.06-1.09 (m, 2H), 1.17-1.27 (m, 2H), 1.42-1.47 (m,10H), 1.88-1.90 (m, 1H), 2.21-2.26 (m, 1H), 2.33-2.39 (m, 1H), 2.61-2.65(m, 1H), 2.95-2.99 (m, 1H), 3.85 (s, 3H), 3.86-3.90 (m, 1H), 3.99-4.00(m, 1H), 4.43-4.45 (m, 1H), 5.13 (d, J=10.8 Hz, 1H), 5.31 (d, J=18.0 Hz,1H), 5.77-5.80 (m, 1H), 5.99 (b, 1H), 7.02 (d, J=9.0 Hz, 2H), 7.51 (t,J=7.3 Hz, 1H), 7.68 (t, J=7.5 Hz, 1H), 7.76 (s, 1H), 7.84 (d, J=8.5 Hz,1H), 8.09 (d, J=8.5 Hz, 2H), 8.15 (d, J=8.0 Hz, 1H);

LC-MS (retention time: 2.02 min, method B), MS m/z 677 (M⁺+H).

Step 3:

This product was prepared by the same method as described in Example 11,Step 5 as a white solid, except using the product of Example 26, Step 2instead.

LC-MS (retention time: 1.53 min, method B), MS m/z 577 (M⁺+H).

Step 4:

Compound 26 was prepared by the same method as described in Example 11,Step 6, except using the product of Example 26, Step 3 instead.

¹H NMR (CD₃OD) δ 0.93-1.09 (m, 12H), 1.26-1.30 (m, 10H), 1.44-1.46 (m,1H), 1.87-1.90 (m, 1H), 2.21-2.26 (m, 1H), 2.36-2.41 (m, 1H), 2.70-2.75(m, 1H), 2.93-2.97 (m, 1H), 3.86 (s, 3H), 4.18-4.25 (m, 1H), 4.30 (b,1H), 4.46-4.48 (m, 1H), 4.55-4.58 (m, 1H), 5.12 (d, J=10.5 Hz, 1H), 5.29(d, J=18.0 Hz, 1H), 5.72-5.79 (m, 1H), 6.08 (b, 1H), 7.02 (d, J=9.0 Hz,2H), 7.44 (t, J=7.3 Hz, 1H), 7.66 (t, J=7.5 Hz, 1H), 7.75 (s, 1H), 7.83(d, J=8.5 Hz, 1H), 8.09 (d, J=8.5 Hz, 2H), 8.15 (d, J=8.0 Hz, 1H);

LC-MS (retention time: 2.03 min, method B), MS m/z 790 (M⁺+H).

Example 27 Preparation of Compound 27

Step 1:

This product was prepared by the same method as described in Example 25,Step 1, except using 4-pyridylboronic acid instead.

¹H NMR (CD₃OD) δ 1.43, 1.46 (rotamers, 9H), 2.53-2.56 (m, 1H), 2.80-2.89(m, 1H), 3.90-3.93 (m, 1H), 4.00-4.05 (m, 1H), 4.50-4.57 (m, 1H), 6.00,6.05 (rotamers, 1H), 7.80 (t, J=7.3 Hz, 1H), 7.87 (t, J=7.5 Hz, 1H),8.08 (d, J=8.5 Hz, 1H), 8.32 (d, J=8.0 Hz, 1H), 8.49 (s, 1H), 8.84 (d,J=6.0 Hz, 2H), 8.84 (d, J=6.5 Hz, 2H);

LC-MS (retention time: 1.39 min, method B), MS m/z 436 (M⁺+H).

Step 2:

The product was prepared by the same method as described in Example 11,Step 4, except using the product of Example 27, Step 1 instead.

¹H NMR (CD₃OD) δ 1.06-1.09 (m, 2H), 1.17-1.27 (m, 2H), 1.42-1.46 (m,10H), 1.88-1.90 (m, 1H), 2.21-2.26 (m, 1H), 2.33-2.39 (m, 1H), 2.61-2.65(m, 1H), 2.95-2.99 (m, 1H), 3.88-3.90 (m, 1H), 4.01-4.08 (m, 1H),4.43-4.45 (m, 1H), 5.15 (d, J=10.8 Hz, 1H), 5.32 (d, J=18.0 Hz, 1H),5.77-5.80 (m, 1H), 6.10 (b, 1H), 7.79 (t, J=7.3 Hz, 1H), 7.88 (t, J=7.5Hz, 1H), 8.08 (d, J=8.5 Hz, 1H), 8.31 (d, J=8.0 Hz, 1H), 8.47 (s, 1H),8.79 (d, J=7.0 Hz, 2H), 8.86 (d, J=6.5 Hz, 2H);

LC-MS (retention time: 1.49 min, method B), MS m/z 648 (M⁺+H).

Step 3:

This product was prepared by the same method as described in Example 11,Step 5 as a white solid, except using the product of Example 27, Step 2instead.

LC-MS (retention time: 0.96 min, method B), MS m/z 548 (M⁺+H).

Step 4:

Compound 27 was prepared by the same method as described in Example 11,Step 6, except using the product of Example 27, Step 3 instead.

¹H NMR (CD₃OD) δ 0.94-1.09 (m, 12H), 1.22-1.26 (m, 10H), 1.44-1.49 (m,1H), 1.88-1.92 (m, 1H), 2.22-2.25 (m, 1H), 2.41-2.44 (m, 1H), 2.70-2.75(m, 1H), 2.93-2.98 (m, 1H), 4.18-4.21 (m, 1H), 4.25 (b, 1H), 4.53-4.62(m, 2H), 5.12 (d, J=10.0 Hz, 1H), 5.29 (d, J=20.0 Hz, 1H), 5.72-5.77 (m,1H), 6.12 (b, 1H), 7.67 (t, J=7.3 Hz, 1H), 7.82 (t, J=7.5 Hz, 1H), 8.02(d, J=8.5 Hz, 1H), 8.29 (d, J=8.0 Hz, 1H), 8.31 (s, 1H), 8.55 (d, J=7.0Hz, 2H), 8.76 (d, J=6.5 Hz, 2H);

LC-MS (retention time: 1.49 min, method B), MS m/z 761 (M⁺+H).

Example 28 Preparation of Compound 28

Step 1:

This product was prepared by the same method as described in Example 25,Step 1, except using 4-N,N-dimethylamino-phenylboronic acid instead.

LC-MS (retention time: 1.64 min, method B), MS m/z 478 (M⁺+H).

Step 2:

This product was prepared by the same method as described in Example 11,Step 4, except using the product of Example 28, Step 1 instead.

LC-MS (retention time: 1.70 min, method B), MS m/z 690 (M⁺+H).

Step 3:

This product was prepared by the same method as described in Example 11,Step 5 as a white solid, except using the product of Example 28, Step 2instead.

LC-MS (retention time: 1.20 min, method B), MS m/z 590 (M⁺+H).

Step 4:

Compound 28 was prepared by the same method as described in Example 11,Step 6, except using the product of Example 28, Step 3 instead.

¹H NMR (d₆-DMSO) δ 0.92-1.10 (m, 13H), 1.30 (s, 9H), 1.35-1.38 (m, 1H),1.68-1.71 (m, 1H), 2.12-3.00 (m, 2H), 2.59-2.62 (m, 1H), 2.91-2.95 (m,1H), 2.99 (s, 6H), 3.93-4.10 (m, 2H), 4.32-4.40 (m, 2H), 5.09 (d, J=11.5Hz, 1H), 5.23 (d, J=19.0 Hz, 1H), 5.54-5.64 (m, 1H), 5.92 (b, 1H), 6.83(d, J=9.0 Hz, 2H), 7.42 (t, J=7.3 Hz, 1H), 7.70 (t, J=7.5 Hz, 1H), 7.81(s, 1H), 7.87 (d, J=8.5 Hz, 1H), 8.04 (d, J=9.0 Hz, 2H), 8.15 (d, J=8.0Hz, 1H);

LC-MS (retention time: 1.72 min, method B), MS m/z 803 (M⁺+H).

Example 29 Preparation of Compound 29

Step 1:

This product was prepared by the same method as described in Example 25,Step 1, except using 4-cyano-phenylboronic acid instead.

LC-MS (retention time: 1.87 min, method B), MS m/z 460 (M⁺+H).

Step 2:

This product was prepared by the same method as described in Example 11,Step 4, except using the product of Example 29, Step 1 instead.

LC-MS (retention time: 1.88 min, method B), MS m/z 672 (M⁺+H).

Step 3:

This product was prepared by the same method as described in Example 11,Step 5 as a white solid, except using the product of Example 29, Step 2instead.

LC-MS (retention time: 1.41 min, method B), MS m/z 572 (M⁺+H).

Step 4:

Compound 29 was prepared by the same method as described in Example 11,Step 6 as a white solid, except using the product of Example 29, Step 3instead.

¹H NMR (CD₃OD) δ 0.92-1.09 (m, 12H), 1.25-1.26 (m, 10H), 1.42-1.46 (m,1H), 1.86-1.89 (m, 1H), 2.20-2.22 (m, 1H), 2.33-2.34 (m, 1H), 2.68-2.71(m, 1H), 2.93-2.95 (m, 1H), 4.13-4.28 (m, 2H), 4.49-4.60 (m, 2H), 5.12(d, J=10.5 Hz, 1H), 5.28 (d, J=18.0 Hz, 1H), 5.71-5.80 (m, 1H), 6.09 (b,1H), 7.56 (t, J=7.3 Hz, 1H), 7.74 (t, J=7.5 Hz, 1H), 7.83 (d, J=10.5 Hz,2H), 7.93 (d, J=7.5 Hz, 1H), 8.01 (s, 1H), 8.22 (d, J=7.5 Hz, 1H), 8.37(d, J=10.5 Hz, 2H);

LC-MS (retention time: 1.87 min, method B), MS m/z 785 (M⁺+H).

Example 30 Preparation of Compound 30

Step 1:

This product was prepared by the same method as described in Example 25,Step 1, except using 3-furanoboronic acid instead.

LC-MS (retention time: 1.85 min, method B), MS m/z 425 (M⁺+H).

Step 2:

This product was prepared by the same method as described in Example 11,Step 4, except using the product of Example 30, Step 1 instead.

LC-MS (retention time: 1.88 min, method B), MS m/z 637 (M⁺+H).

Step 3:

This product was prepared by the same method as described in Example 11,Step 5, except using the product of Example 30, Step 2 instead.

LC-MS (retention time: 1.38 min, method B), MS m/z 537 (M⁺+H).

Step 4:

Compound 30 was prepared by the same method as described in Example 11,Step 6 as a white solid, except using the product of Example 30, Step 3instead.

¹H NMR (CD₃OD) δ 0.95-1.09 (m, 12H), 1.23-1.30 (m, 10H), 1.43-1.46 (m,1H), 1.87-1.90 (m, 1H), 2.21-2.23 (m, 1H), 2.30-2.34 (m, 1H), 2.64-2.70(m, 1H), 2.93-2.96 (m, 1H), 4.11-4.29 (m, 2H), 4.41-4.44 (m, 1H),4.54-4.56 (m, 1H), 5.12 (d, J=10.5 Hz, 1H), 5.29 (d, J=17.5 Hz, 1H),5.71-5.80 (m, 1H), 6.02 (b, 1H), 7.00 (s, 1H), 7.44 (t, J=7.2 Hz, 1H),7.52 (s, 1H), 7.57 (s, 1H), 7.66 (t, J=7.0 Hz, 1H), 7.79 (d, J=8.0 Hz,1H), 8.14-8.17 (m, 2H);

LC-MS (retention time: 1.93 min, method B), MS m/z 750 (M⁺+H).

Example 31 Preparation of Compound 31

Step 1:

This product was prepared by the same method as described in making ofExample 21, Step 2 as a white solid, except using 3-bromo-isoquinoline(Atkins et al, JOC, 1973, 400) instead.

¹H NMR (CDCl₃) δ 7.60-7.62 (m, 2H), 7.71-7.73 (m, 2H), 8.12 (s, 1H),8.99 (s, 1H);

LC-MS (retention time: 0.78 min, method B), MS m/z 224, 226 (M⁺+H).

Step 2:

This product was prepared by the same method as described in Example 11,Step 2 as white solid, except using 3-bromo-isoquinoline 2-oxideinstead.

¹H NMR (CDCl₃) δ 7.66-7.71 (m, 1H), 7.74-7.76 (m, 2H), 7.83 (s, 1H),8.29 (d, J=8.5 Hz, 1H);

LC-MS (retention time: 1.55 min, method B), MS m/z 242, 244 (M⁺+H).

Step 3:

This product was prepared by the same method as described in Example 11,Step 3 as a foam, except using 3-bromo-1-chloro-isoquinoline instead.

¹H NMR (CD₃OD) δ 1.43, 1.44 (rotamers, 9H), 2.41-2.47 (m, 1H), 2.69-2.72(m, 1H), 3.80-3.84 (m, 1H), 3.88-3.90 (m, 1H), 4.46-4.52 (m, 1H), 5.76(b, 1H), 7.57-7.61 (m, 2H), 7.73-7.75 (m, 2H), 8.15 (d, J=8.0 Hz, 1H);

LC-MS (retention time: 1.79 min, method B), MS m/z 437, 439 (M⁺+H).

Step 4:

A mixture of 2-tributylstannanyl-pyrazine (44 mg, 0.12 mmol),tetrakis(triphenylphosphine) palladium (0) (12 mg, 0.01 mmol) and theproduct of Example 31, Step 3 (44 mg, 0.1 mmol) in toluene (1 mL) washeated to reflux for 3 h. After removing the volatiles in vacuo, theresidue was purified by prep-HPLC to yield 35 mg (80%) of the desiredproduct as a yellow solid.

LC-MS (retention time: 1.77 min, method B), MS m/z 437 (M⁺+H).

Step 5:

This product was prepared by the same method as described in Example 11,Step 4, except using the product of Example 31, Step 4 instead.

LC-MS (retention time: 1.78 min, method B), MS m/z 649 (M⁺+H).

Step 6:

This product was prepared by the same method as described in Example 11,Step 5 as a white solid, except using the product of Example 31, Step 5instead.

LC-MS (retention time: 1.26 min, method B), MS m/z 549 (M⁺+H).

Step 7:

Compound 31 was prepared by the same method as described in Example 11,Step 6, except using the product of Example 31, Step 6 instead.

¹H NMR (CD₃OD) δ 0.95-1.10 (m, 12H), 1.24-1.27 (m, 10H), 1.44-1.47 (m,1H), 1.87-1.90 (m, 1H), 2.19-2.22 (m, 1H), 2.38-2.44 (m, 1H), 2.71-2.76(m, 1H), 2.93-2.96 (m, 1H), 4.18-4.28 (m, 2H), 4.50-4.61 (m, 2H), 5.12(d, J=10.5 Hz, 1H), 5.29 (d, J=17.5 Hz, 1H), 5.71-5.80 (m, 1H), 6.12 (b,1H), 7.60 (t, J=7.2 Hz, 1H), 7.77 (t, J=7.0 Hz, 1H), 7.97 (d, J=8.5 Hz,1H), 8.26 (d, J=8.5 Hz, 1H), 8.44 (s, 1H), 8.59 (s, 1H), 8.70 (s, 1H),9.61 (s, 1H);

LC-MS (retention time: 1.84 min, method B), MS m/z 762 (M⁺+H).

Example 32 Preparation of Compound 32

Step 1:

This product, 2-oxy-isoquinoline-3-carbonitrile, was prepared by thesame method as described in Example 21, Step 2 as a white solid, exceptusing 3-cyano-isoquinoline instead.

¹H NMR (DMSO-d₆) δ 7.74 (t, J=8.0 Hz, 1H), 7.84 (t, J=8.2 Hz, 1H), 7.97(d, J=8.5 Hz, 1H), 8.03 (d, J=8.5 Hz, 1H), 8.85 (s, 1H), 9.17 (s, 1H);

LC-MS (retention time: 0.48 min, method B), MS m/z 171 (M⁺+H).

Step 2:

This product, 1-chloro-isoquinoline-3-carbonitrile was prepared by thesame method as described in Example 11, Step 2 as white solid, exceptusing 3-cyano-isoquinoline 2-oxide instead.

¹H NMR (CDCl₃) δ 7.87-7.91 (m, 2H), 7.92-7.94 (m, 1H), 8.09 (s, 1H),8.42-8.44 (m, 1H);

LC-MS (retention time: 1.22 min, method B), MS m/z 189 (M⁺+H).

Step 3:

This product was prepared by the same method as described in Example 1,Step 5, except using 1-chloro-isoquinoline-3-carbonitrile instead.

¹HNMR (CD₃OD) δ 1.05 (s, 9H), 1.17 (s, 9H), 2.34-2.40 (m, 1H), 2.71-2.78(m, 1H), 4.09-4.11 (m, 1H), 4.21 (b, 1H), 4.48-4.52 (m, 1H), 4.68-4.72(m, 1H), 5.89 (b, 1H), 7.74 (t, J=7.5 Hz, 1H), 7.86 (t, J=7.5 Hz, 1H),7.94-7.97 (m, 2H), 8.31 (d, J=8.0 Hz, 1H);

LC-MS (retention time: 1.66 min, method B), MS m/z 497 (M⁺+H).

Step 4:

Compound 32 was prepared by the same method as described in Example 1,Step 9 as a white solid, except using the product of Example 32, Step 3instead.

¹H NMR (CD₃OD) δ 1.04-1.09 (m, 12H), 1.20-1.27 (m, 10H), 1.39-1.45 (m,1H), 1.85-1.88 (m, 1H), 2.20-2.30 (m, 2H), 2.63-2.71 (m, 1H), 2.91-2.97(m, 1H), 4.09-4.13 (m, 1H), 4.23 (d, J=9.3 Hz, 1H), 4.49-4.58 (m, 2H),5.13 (d, J=10.5 Hz, 1H), 5.28 (d, J=18.0 Hz, 1H), 5.69-5.81 (m, 1H),5.92 (b, 1H), 6.60 (d, J=10.0 Hz, 1H), 7.72 (t, J=7.5 Hz, 1H), 7.86 (t,J=7.5 Hz, 1H), 7.96-7.99 (m, 2H), 8.29 (d, J=8.0 Hz, 1H);

LC-MS (retention time: 1.75 min, method B), MS m/z 714 (M⁺+H).

Example 33 Preparation of Compound 33

Step 1:

This product, 3-methyl-isoquinoline 2-oxide, was prepared by the samemethod as described in Example 21, Step 2 as a white solid, except using3-methyl-isoquinoline instead.

¹H NMR (CD₃OD) δ 2.64 (s, 3H), 7.64-7.72 (m, 2H), 7.88-7.95 (m, 2H),9.05 (s, 1H);

LC-MS (retention time: 0.61 min, method B), MS m/z 160 (M⁺+H).

Step 2:

This product, 1-chloro-3-methyl-isoquinoline was prepared by the samemethod as described in Example 11, Step 2 as white solid, except using3-methyl-isoquinoline 2-oxide instead.

¹H NMR (CDCl₃) δ 2.65 (s, 3H), 7.25 (s, 1H), 7.61 (t, J=7.5 Hz, 1H),7.69 (t, J=7.5 Hz, 1H), 7.74 (d, J=8.0 Hz, 1H), 8.27 (d, J=8.5 Hz, 1H);

LC-MS (retention time: 1.47 min, method B), MS m/z 178 (M⁺+H).

Step 3:

This product was prepared by the same method as described in Example 1,Step 5 as a white solid, except using 1-chloro-3-methyl-isoquinolineinstead.

¹HNMR (CD₃OD) δ 1.05 (s, 9H), 1.23 (s, 9H), 2.51 (s, 3H), 2.34-2.40 (m,1H), 2.72-2.78 (m, 1H), 4.05-4.12 (m, 1H), 4.26 (b, 1H), 4.41 (d, J=10Hz, 1H), 4.62-4.67 (m, 1H), 5.90 (b, 1H), 7.14 (s, 1H), 7.38 (t, J=7.5Hz, 1H), 7.62 (t, J=7.5 Hz, 1H), 7.68 (d, J=8.0 Hz, 1H), 8.14 (d, J=8.0Hz, 1H);

LC-MS (retention time: 1.84 min, method B), MS m/z 486 (M⁺+H).

Step 4:

Compound 33 was prepared by the same method as described in Example 1,Step 9 as a white solid, except using the product of Example 33, Step 3instead.

¹H NMR (CD₃OD) δ 0.99-1.09 (m, 12H), 1.23-1.25 (m, 10H), 1.41-1.45 (m,1H), 1.86-1.90 (m, 1H), 2.21-2.31 (m, 2H), 2.52 (s, 3H), 2.58-2.61 (m,1H), 2.91-2.97 (m, 1H), 4.08-4.12 (m, 1H), 4.28 (b, 1H), 4.40 (d, J=10.0Hz, 1H), 4.50-4.55 (m, 1H), 5.12 (d, J=10.0 Hz, 1H), 5.30 (d, J=18.0 Hz,1H), 5.71-5.81 (m, 1H), 5.93 (b, 1H), 7.13 (s, 1H), 7.38 (t, J=7.5 Hz,1H), 7.62 (t, J=7.5 Hz, 1H), 7.68 (d, J=8.0 Hz, 1H), 8.12 (d, J=8.0 Hz,1H), 9.12 (b, 1H);

LC-MS (retention time: 1.85 min, method B), MS m/z 698 (M⁺+H).

Example 34 Preparation of Compound 34

Step 1:

This product, 3-cyclopropyl-isoquinoline 2-oxide was prepared by thesame method as described in Example 21, Step 2 as a white solid, exceptusing 3-cyclopropyl-isoquinoline (L. Flippin, J. Muchowski, J. O. C,1993, 2631-2632) instead.

LC-MS (retention time: 0.95 min, method B), MS m/z 186 (M⁺+H).

Step 2:

This product, 1-chloro-3-cyclopropyl-isoquinoline was prepared by thesame method as described in Example 11, Step 2 as white solid, exceptusing 3-cyclopropyl-isoquinoline 2-oxide instead.

¹H NMR (CD₃OD) δ 1.00-1.04 (m, 4H), 2.11-2.18 (m, 1H), 7.55 (s, 1H),7.61 (t, J=8.0 Hz, 1H), 7.72 (t, J=8.0 Hz, 1H), 7.83 (d, J=13.5 Hz, 1H),8.27 (d, J=14.5 Hz, 1H);

LC-MS (retention time: 1.70 min, method B), MS m/z 204 (M⁺+H).

Step 3:

This product was prepared by the same method as described in Example 1,Step 5 as a white solid, except using1-chloro-3-cyclopropyl-isoquinoline instead.

¹HNMR (CD₃OD) δ 0.93-1.05 (m, 13H), 1.29 (s, 9H), 2.06-2.10 (m, 1H),2.39-2.44 (m, 1H), 2.70-2.76 (m, 1H), 4.05-4.12 (m, 1H), 4.27 (b, 1H),4.35 (d, J=10.0 Hz, 1H), 4.62-4.67 (m, 1H), 5.78 (b, 1H), 7.18 (s, 1H),7.38 (t, J=7.5 Hz, 1H), 7.61 (t, J=7.5 Hz, 1H), 7.66 (d, J=8.0 Hz, 1H),8.09 (d, J=8.0 Hz, 1H);

LC-MS (retention time: 1.96 min, method B), MS m/z 512 (M⁺+H).

Step 4:

Compound 34 was prepared by the same method as described in Example 1,Step 9 as a white solid, except using the product of Example 34, Step 3instead.

¹HNMR (CD₃OD) δ 0.93-1.09 (m, 16H), 1.24-1.30 (m, 10H), 1.42-1.46 (m,1H), 1.87-1.90 (m, 1H), 2.06-2.11 (m, 1H), 2.21-2.32 (m, 2H), 2.56-2.61(m, 1H), 2.92-2.97 (m, 1H), 4.08-4.12 (m, 1H), 4.28 (b, 1H), 4.32 (d,J=10.0 Hz, 1H), 4.48-4.53 (m, 1H), 5.12 (d, J=10.5 Hz, 1H), 5.30 (d,J=17.5 Hz, 1H), 5.72-5.77 (m, 1H), 5.82 (b, 1H), 7.18 (s, 1H), 7.36 (t,J=7.5 Hz, 1H), 7.60 (t, J=7.5 Hz, 1H), 7.67 (d, J=8.0 Hz, 1H), 8.07 (d,J=8.0 Hz, 1H);

LC-MS (retention time: 2.00 min, method B), MS m/z 724 (M⁺+H).

Example 35 Preparation of Compound 35

Step 1:

A mixture of 3-hydroxy-isoquinoline (725 mg, 5.0 mmol), cesium carbonate(4.89 g, 15.0 mmol), MeI (781 mg, 5.5 mmol) in DMF (50 mL) was stirredat the ambient temperature for 12 h. The mixture was diluted with EtOAc(200 mL), filtered, washed with water (200 mL, ×2) and 1M NaOH (aq),brine respectively. The organic layer was dried over MgSO₄, filtered,evaporated. The residue was purified by prep-HPLC to yield 120 mg (15%)of the desired product as a white solid.

¹H NMR (CDCl₃) δ 4.03 (s, 3H), 6.99 (s, 1H), 7.36 (t, J=8.0 Hz, 1H),7.56 (t, J=8.2 Hz, 1H), 7.68 (d, J=8.5 Hz, 1H), 7.87 (J=8.5 Hz, 1H);

LC-MS (retention time: 0.54 min, method B), MS m/z 160 (M⁺+H).

Step 2:

This product, 3-methoxy-isoquinoline 2-oxide, was prepared by the samemethod as described in Example 21, Step 2 as a white solid, except using3-methoxy-isoquinoline instead.

LC-MS (retention time: 0.83 min, method B), MS m/z 176 (M⁺+H).

Step 3:

This product, 1-chloro-3-methoxy-isoquinoline was prepared by the samemethod as described in Example 11, Step 2 as white solid, except using3-methoxy-isoquinoline 2-oxide instead.

LC-MS (retention time: 1.62 min, method B), MS m/z 194 (M⁺+H).

Step 4:

This product was prepared by the same method as described in Example 1,Step 5 as a white solid, except using 1-chloro-3-methoxy-isoquinolineinstead.

¹HNMR (CD₃OD) δ 1.05 (s, 9H), 1.23 (s, 9H), 2.35-2.43 (m, 1H), 2.72-2.79(m, 1H), 3.96 (s, 3H), 4.01-4.11 (m, 1H), 4.26 (b, 1H), 4.48 (d, J=10.0Hz, 1H), 4.62-4.67 (m, 1H), 5.83 (b, 1H), 6.61 (s, 1H), 7.25 (t, J=7.5Hz, 1H), 7.54 (t, J=7.5 Hz, 1H), 7.63 (d, J=8.1 Hz, 1H), 8.08 (d, J=8.4Hz, 1H);

LC-MS (retention time: 1.82 min, method B), MS m/z 502 (M⁺+H).

Step 5:

Compound 35 was prepared by the same method as described in Example 1,Step 9 as a white solid, except using the product of Example 35, Step 4instead.

¹H NMR (CD₃OD) δ 1.04-1.08 (m, 12H), 1.24-1.27 (m, 10H), 1.43-1.45 (m,1H), 1.86-1.89 (m, 1H), 2.21-2.26 (m, 1H), 2.30-2.34 (m, 1H), 2.62-2.66(m, 1H), 2.91-2.97 (m, 1H), 3.99 (s, 3H), 4.09-4.12 (m, 1H), 4.27-4.28(m, 1H), 4.46 (d, J=10.0 Hz, 1H), 4.51-4.58 (m, 1H), 5.12 (d, J=10.5 Hz,1H), 5.30 (d, J=18.0 Hz, 1H), 5.72-5.76 (m, 1H), 5.88 (b, 1H), 6.62 (s,1H), 7.26 (t, J=7.5 Hz, 1H), 7.55 (t, J=7.5 Hz, 1H), 7.65 (d, J=8.0 Hz,1H), 8.06 (d, J=8.5 Hz, 1H);

LC-MS (retention time: 1.85 min, method B), MS m/z 714 (M⁺+H).

Example 36 Preparation of Compound 36

Step 1:

A mixture of 4-methoxy-2-methyl-benzoic acid (5.00 g, 30.1 mmol) andthionyl chloride (20.0 g, 0.17 mol) was heated to reflux for 30 min.Removed the volatile in vacuo. After pumping overnight, the viscous oilyacid chloride was used as crude for the next reaction without anypurification.

To a solution of 4-methoxy-2-methyl-benzoyl chloride in CH₂Cl₂ (60 mL)at 0° C. was added diethylamine dropwise. The formed mixture was allowedto warm up to the ambient temperature for 2 h with stirring. Removed thevolatiles in vacuo. The residue was triturated with EtOAc (100 mL) andfiltered. The filtrate was washed with 1M HCl, 1M NaOH and brine, driedover MgSO₄. Evaporation of the solvent yielded 6.51 g (98%) of thedesired product as a viscous oil.

LC-MS (retention time: 1.20 min, method B), MS m/z 222 (M⁺+H).

Step 2:

To a solution of N,N-diethyl-4-methoxy-2-methyl-benzamide (221 mg, 1.0mmol) in THF (2 mL) at −78° C. was added n-BuLi (0.84 mL of 2.5 M inhexane, 2.10 mmol) dropwise. The formed orange solution was kept at thistemperature for additional 30 min before dropwise addition ofbenzonitrile (103 mg, 1.0 mmol). The final solution was allowed to warmup to the ambient temperature over night with stirring. Quenched withiced 5% citric acid. Filtered, washed with water, dried. Triturationwith 2:1 hexane-EtOAc (5 mL) yielded 205 mg (82%) of the desired productas a white solid.

¹H NMR (d₆-DMSO) δ 3.89 (s, 3H), 6.84 (s, 1H), 7.05-7.07 (m, 1H), 7.18(d, J=2.5 Hz, 1H), 7.44-7.51 (m, 3H), 7.78 (d, J=7.0 Hz, 1H), 8.11 (d,J=9.0 Hz, 1H);

LC-MS (retention time: 1.20 min, method B), MS m/z 252 (M⁺+H).

Step 3:

This product, 1-chloro-6-methoxy-3-phenyl-isoquinoline, was prepared bythe same method as described in Example 11, Step 2 as a white solid,except using 6-methoxy-3-phenyl-2H-isoquinolin-1-one instead.

¹H NMR (CDCl₃) δ 3.97 (s, 3H), 7.12 (d, J=2.5 Hz, 1H), 7.23-7.26 (m,1H), 7.40-7.42 (m, 1H), 7.46-7.50 (m, 2H), 7.89 (s, 1H), 8.08 (d, J=7.0Hz, 2H), 8.21 (d, J=9.0 Hz, 1H);

LC-MS (retention time: 1.90 min, method B), MS m/z 270, 271 (M⁺+H).

Step 4:

To a solution of the product of Example 21, Step 1 (320 mg, 0.57 mmol)in DMSO (5 mL) was added potassium tert-butoxide (321 mg, 2.87 mmol).The formed solution was stirred at the ambient temperature for 30 minbefore addition of 1-chloro-6-methoxy-3-phenyl-isoquinoline (Example 36,Step 3) (155 mg, 0.57 mmol). The final solution was stirred for 12 h.Quenched with iced water, acidified with 1M HCl to pH 4, extracted withEtOAc (20 mL, ×2). The organic layers were washed with brine, dried overMgSO₄, filtered, evaporated. The residue was purified by prep-HPLC (40%B to 100% B, 15 min gradient) to yield 289 mg (64%) of Compound 36 as awhite solid.

¹H NMR (CD₃OD) δ 0.95-1.05 (m, 12H), 1.24-1.32 (m, 10H), 1.44-1.46 (m,1H), 1.87-1.90 (m, 1H), 2.20-2.26 (m, 1H), 2.30-2.36 (m 1H), 2.65-2.71(m, 1H), 2.93-2.97 (m, 1H), 3.94 (s, 3H), 4.12-4.28 (m, 2H), 4.38-4.52(m, 2H), 5.12 (d, J=10.0 Hz, 1H), 5.28 (d, J=17.0 Hz, 1H), 5.69-5.74 (m,1H), 6.05 (b, 1H), 7.06-7.07 (m, 1H), 7.26 (s, 1H), 7.37-7.39 (m, 1H),7.44-7.48 (m, 2H), 7.77 (s, 1H), 8.07 (d, J=9.0 Hz, 1H), 8.15 (d, J=8.5Hz, 2H);

LC-MS (retention time: 2.02 min, method B), MS m/z 790 (M⁺+H).

Example 37 Preparation of Compound 37

Step 1:

To a solution of N,N-diethyl-4-methoxy-2-methyl-benzamide (633 mg, 2.9mmol) in THF (15 mL) at −78° C. was added n-BuLi (2.3 mL of 2.5 M inhexane, 5.74 mmol) dropwise. The formed red solution was kept at thistemperature for additional min before being cannulated to a solution ofthiazole-2-carboxylic acid ethyl ester (A. Medici et al, TetrahedronLett. 1983, p 2901) (450 mg, 2.9 mmol) in THF (5 mL) at −78° C. Thefinal dark green solution was kept to this temperature for 2 h withstirring. Quenched with sat. NH₄Cl (aq) and extracted with EtOAc (50mL). The organic layer was washed with sat. NH₄Cl (aq) and brine, dried,purified by flash column chromatography, eluting with 2:1 EtOAc:hexaneto provide 405 mg (45%) of the desired product as an off-white viscousoil.

¹H NMR (CDCl₃) δ 1.08 (t, J=7.0 Hz, 6H), 3.22 (b, 2H), 3.44 (b, 2H),3.79 (s, 3H), 4.59 (s, 2H), 6.79-6.81 (m, 1H), 6.86 (d, J=2.5 Hz, 1H),7.16 (d, J=8.5 Hz, 1H), 7.66 (d, J=3.0 Hz, 1H), 8.00 (d, J=3.0 Hz, 1H);

LC-MS (retention time: 1.30 min, method B), MS m/z 333 (M⁺+H).

Step 2:

A mixture ofN,N-diethyl-4-methoxy-2-(2-oxo-2-thiazol-2-yl-ethyl)-benzamide (405 mg,1.22 mmol) and NH₄OAc (3.0 g, 38.9 mmol) was heated to 140° C. in asealed tube for 1 h. The melted solution was poured into iced water,filtered, washed the cake thoroughly with water. The dried brownishsolid (240 mg, 76%) was used as crude for the next reaction withoutfurther purification.

LC-MS (retention time: 1.24 min, method B), MS m/z 259 (M⁺+H).

Step 3:

This product, 1-chloro-6-methoxy-3-thiazol-2-yl-isoquinoline, wasprepared by the same method as described in Example 11, Step 2 as awhite solid, except using 6-methoxy-3-thiazol-2-yl-2H-isoquinolin-1-oneinstead.

¹H NMR (CDCl₃) δ 3.97 (s, 3H), 7.16 (d, J=4.0 Hz, 1H), 7.27-7.31 (m,1H), 7.46 (d, J=5.0 Hz, 1H), 7.93 (d, J=5.5 Hz, 1H), 8.22 (d, J=15.5 Hz,1H), 8.39 (s, 1H);

LC-MS (retention time: 1.66 min, method B), MS m/z 277 (M⁺+H).

Step 4:

Compound 37 was prepared by the same method as described in Example 36,Step 4, except using 1-chloro-6-methoxy-3-thiazol-2-yl-isoquinolineinstead.

¹H NMR (CD₃OD) δ 0.97-1.09 (m, 12H), 1.24-1.29 (m, 10H), 1.44-1.46 (m,1H), 1.87-1.90 (m, 1H), 2.20-2.26 (m, 1H), 2.30-2.36 (m, 1H), 2.65-2.71(m, 1H), 2.93-2.96 (m, 1H), 3.96 (s, 3H), 4.12-4.27 (m, 2H), 4.38-4.52(m, 2H), 5.12 (d, J=10.5 Hz, 1H), 5.29 (d, J=17.5 Hz, 1H), 5.69-5.74 (m,1H), 5.99 (b, 1H), 7.14 (d, J=9.0 Hz, 1H), 7.33 (s, 1H), 7.66 (d, J=3.5Hz, 1H), 7.93 (d, J=3.0 Hz, 1H), 8.05 (s, 1H), 8.11 (d, J=9.0 Hz, 1H),9.14 (b, 1H);

LC-MS (retention time: 1.89 min, method B), MS m/z 797 (M⁺+H).

Example 38 Preparation of Compound 38

Step 1:

A mixture of 3-hydroxy-isoxazole-5-carboxylic acid methyl ester (5.72 g,0.04 mol), methyl iodide (6.82 g, 0.044 mol) and cesium carbonate (39.1g, 0.12 mol) in DMF (200 mL) was stirred at the ambient temperature overnight. Diluted with EtOAc (1 L), filtered. The filtrate was washed withwater (1 L, ×2), 1M NaOH and brine respectively, dried over MgSO₄,evaporated in vacuo to afford 4.80 g (76%) of the desired product as awhite solid. The product obtained here was used as crude without furtherpurification.

¹H NMR (CDCl₃) δ 3.92 (s, 3H), 4.00 (s, 3H), 6.51 (s, 1H);

LC-MS (retention time: 0.69 min, method B), MS m/z 158 (M⁺+H).

Step 2:

This product,N,N-diethyl-4-methoxy-2-[2-(3-methoxy-isoxazol-5-yl)-2-oxo-ethyl]-benzamide,was prepared by the same method as described in Example 37, Step 1,except using 3-methoxy-isoxazole-5-carboxylic acid methyl ester instead.

LC-MS (retention time: 1.28 min, method B), MS m/z 347 (M⁺+H).

Step 3:

This product,6-methoxy-3-(3-methoxy-isoxazol-5-yl)-2H-isoquinolin-1-one, was preparedby the same method as described in Example 37, Step 2, except usingN,N-diethyl-4-methoxy-2-[2-(3-methoxy-isoxazol-5-yl)-2-oxo-ethyl]-benzamideinstead.

¹H NMR (DMSO-d₆) δ 3.89 (s, 3H), 3.97 (s, 3H), 7.01 (s, 1H), 7.14-7.16(m, 2H), 7.43 (s, 1H), 8.13 (d, J=8.5 Hz, 1H);

LC-MS (retention time: 1.31 min, method B), MS m/z 273 (M⁺+H).

Step 4:

This product,1-chloro-6-methoxy-3-(3-methoxy-isoxazol-5-yl)-isoquinoline, wasprepared by the same method as described in Example 11, Step 2 as awhite solid, except using6-methoxy-3-(3-methoxy-isoxazole-5-yl)-2H-isoquinolin-1-one instead.

¹H NMR (CDCl₃) δ 3.97 (s, 3H), 4.04 (s, 3H), 6.60 (s, 1H), 7.17 (d,J=2.5 Hz, 1H), 7.31-7.33 (m, 1H), 8.02 (s, 1H), 8.23 (d, J=9.0 Hz, 1H);

LC-MS (retention time: 1.73 min, method B), MS m/z 291, 293 (M⁺+H).

Step 5:

Compound 38 was prepared by the same method as described in Example 36,Step 4, except using1-chloro-6-methoxy-3-(3-methoxy-isoxazole-5-yl)-isoquinoline instead.

¹H NMR (CD₃OD) δ 0.99-1.09 (m, 12H), 1.23-1.28 (m, 10H), 1.44-1.46 (m,1H), 1.87-1.90 (m, 1H), 2.20-2.26 (m, 1H), 2.30-2.36 (m, 1H), 2.65-2.71(m, 1H), 2.93-2.96 (m, 1H), 3.95 (s, 3H), 4.02 (s, 3H), 4.13-4.14 (m,1H), 4.24-4.26 (m, 1H), 4.41-4.42 (m, 1H), 4.52-4.55 (m, 1H), 5.12 (d,J=10.5 Hz, 1H), 5.29 (d, J=17.0 Hz, 1H), 5.72-5.79 (m, 1H), 5.96 (b,1H), 6.60 (s, 1H), 7.15-7.17 (m, 1H), 7.32 (s, 1H), 7.80 (s, 1H), 8.10(d, J=9.0 Hz, 1H);

LC-MS (retention time: 1.95 min, method B), MS m/z 811 (M⁺+H).

Example 39 Preparation of Compound 39

Step 1:

This product,N,N-diethyl-4-methoxy-2-[2-(5-methoxy-oxazol-2-yl)-2-oxo-ethyl]-benzamide,was prepared by the same method as described in Example 37, Step 1,except using 5-methoxy-oxazole-2-carboxylic acid ethyl ester instead.

LC-MS (retention time: 1.24 min, method B), MS m/z 347 (M⁺+H).

Step 2:

This product, 6-methoxy-3-(5-methoxy-oxazol-2-yl)-2H-isoquinolin-1-one,was prepared by the same method as described in Example 37, Step 2,except usingN,N-diethyl-4-methoxy-2-[2-(5-methoxy-oxazol-2-yl)-2-oxo-ethyl]-benzamideinstead.

¹H NMR (DMSO-d₆) δ 3.94 (s, 3H), 4.01 (s, 3H), 6.34 (s, 1H), 6.99 (d,J=2.0 Hz, 1H), 7.12-7.14 (m, 1H), 7.25 (s, 1H), 8.32 (d, J=9.0 Hz, 1H);

LC-MS (retention time: 1.22 min, method B), MS m/z 274 (M⁺+H).

Step 3:

This product, 1-chloro-6-methoxy-3-(5-methoxy-oxazol-2-yl)-isoquinoline,was prepared by the same method as described in Example 11, Step 2 as awhite solid, except using6-methoxy-3-(5-methoxy-oxazole-2-yl)-2H-isoquinolin-1-one instead.

¹H NMR (CDCl₃) δ 3.96 (s, 3H), 4.00 (s, 3H), 6.34 (s, 1H), 7.12 (d,J=2.5 Hz, 1H), 7.28-7.31 (m, 1H), 8.13 (s, 1H), 8.23 (d, J=9.0 Hz, 1H);

LC-MS (retention time: 1.58 min, method B), MS m/z 291, 293 (M⁺+H).

Step 4:

Compound 39 was prepared by the same method as described in Example 36,Step 4, except using1-chloro-6-methoxy-3-(3-methoxy-isoxazole-5-yl)-isoquinoline instead.

¹H NMR (CD₃OD) δ 0.99-1.09 (m, 12H), 1.23-1.28 (m, 10H), 1.44-1.46 (m,1H), 1.87-1.90 (m, 1H), 2.20-2.26 (m, 1H), 2.30-2.36 (m, 1H), 2.65-2.71(m, 1H), 2.93-2.96 (m, 1H), 3.95 (s, 3H), 4.02 (s, 3H), 4.13-4.14 (m,1H), 4.25 (b, 1H), 4.41-4.42 (m, 1H), 4.52-4.55 (m, 1H), 5.12 (d, J=10.0Hz, 1H), 5.29 (d, J=17.0 Hz, 1H), 5.72-5.79 (m, 1H), 6.07 (b, 1H), 6.45(s, 1H), 7.15-7.16 (m, 1H), 7.29 (s, 1H), 7.85 (s, 1H), 8.10 (d, J=9.0Hz, 1H), 9.11 (b, 1H);

LC-MS (retention time: 1.75 min, method B), MS m/z 811 (M⁺+H).

Example 40 Preparation of Compound 40

Step 1:

This product, 1-chloro-6-methoxy-isoquinoline 2-oxide was prepared bythe same method as described in Example 21, Step 2, except using1-chloro-6-methoxy-isoquinoline (the product of Example 11, Step 2)instead.

¹H NMR (CDCl₃) δ 4.00 (s, 3H), 7.14 (d, J=2.5 Hz, 1H), 7.41-7.43 (m,1H), 7.62 (d, J=7.0 Hz, 1H), 8.15 (d, J=9.5 Hz, 1H), 8.36 (d, J=7.0 Hz,1H);

LC-MS (retention time: 0.85 min, method B), MS m/z 210 (M⁺+H).

Step 2:

This product, 1,3-dichloro-6-methoxy-isoquinoline was prepared by thesame method as described in Example 11, Step 2, except using1-chloro-6-methoxy-isoquinoline 2-oxide instead.

¹H NMR (CDCl₃) δ 3.94 (s, 3H), 6.98 (s, 1H), 7.25-7.26 (m, 1H), 7.52 9s,1H), 8.16 (d, J=9.5 Hz, 1H);

LC-MS (retention time: 1.54 min, method B), MS m/z 228, 230 (M⁺+H).

Step 3:

This product was prepared by the same method as described in Example 24,Step 1 as a foam, except using 1,3-dichloro-6-methoxy-isoquinolineinstead.

¹H NMR (CD₃OD) δ 1.43, 1.44 (rotamers, 9H), 2.39-2.44 (m, 1H), 2.68-2.72(m, 1H), 3.80-3.90 (m, 2H), 3.91 (s, 3H), 4.79-4.82 (m, 1H), 5.71 (b,1H), 7.10-7.14 (m, 2H), 7.26 (s, 1H), 7.99-8.01 (m, 1H);

LC-MS (retention time: 1.79 min method B), MS m/z 422 (M⁺+H).

Step 4:

This product was prepared by the same method as described in Example 11,Step 4, except using the product of Example 40, Step 3 instead.

LC-MS (retention time: 1.83 min, method B), MS m/z 635 (M⁺+H).

Step 5:

This product was prepared by the same method as described in Example 11,Step 5 as a white solid, except using the product of Example 40, Step 4instead.

LC-MS (retention time: 1.36 min, method B), MS m/z 535 (M⁺+H).

Step 6:

Compound 40 was prepared by the same method as described in Example 11,Step 6 as a white solid, except using the product of Example 40, Step 5instead.

¹H NMR (CD₃OD) δ 1.07-1.11 (m, 12H), 1.26-1.30 (m, 10H), 1.46-1.48 (m,1H), 1.87-1.91 (m, 1H), 2.21-2.34 (m, 2H), 2.62-2.66 (m, 1H), 2.94-2.99(m, 1H), 3.95 (s, 3H), 4.06-4.11 (m, 1H), 4.26-4.28 (m, 1H), 4.46-4.56(m, 2H), 5.15 (d, J=10.0 Hz, 1H), 5.29 (d, J=17.0 Hz, 1H), 5.72-5.79 (m,1H), 5.89 (b, 1H), 6.63 (d, J=9.0 Hz, 1H), 7.08-7.09 (m, 1H), 7.18 (s,1H), 7.34 (s, 1H), 8.08 (d, J=9.5 Hz, 1H);

LC-MS (retention time: 1.99 min, method B), MS m/z 748 (M⁺+H).

Example 41 Preparation of Compound 41

Step 1:

This product was prepared by the same method as described in Example 30,Step 1, except using the product of Example 40, Step 3 instead.

LC-MS (retention time: 1.85 min, method B), MS m/z 455 (M⁺+H).

Step 2:

This product was prepared by the same method as described in Example 11,Step 4 as a foam, except using the product of Example 41, Step 1instead.

LC-MS (retention time: 1.88 min, method B), MS m/z 667 (M⁺+H).

Step 3:

This product was prepared by the same method as described in Example 11,Step 5 as a white solid, except using the product of Example 41, Step 2instead.

LC-MS (retention time: 1.38 min, method B), MS m/z 567 (M⁺+H).

Step 4:

Compound 41 was prepared by the same method as described in Example 11,Step 6 as a white solid, except using the product of Example 41, Step 3instead.

¹H NMR (CD₃OD) δ 0.99-1.04 (m, 12H), 1.22-1.31 (m, 10H), 1.43-1.45 (m,1H), 1.87-1.89 (m, 1H), 2.22-2.24 (m, 1H), 2.30-2.34 (m, 1H), 2.65-2.68(m, 1H), 2.93-2.96 (m, 1H), 3.92 (s, 3H), 4.11-4.14 (m, 1H), 4.28-4.30(m, 1H), 4.38-4.42 (m, 1H), 4.53-4.55 (m, 1H), 5.12 (d, J=10.0 Hz, 1H),5.29 (d, J=18.0 Hz, 1H), 5.72-5.77 (m, 1H), 5.99 (b, 1H), 6.61 (d, J=5.0Hz, 1H), 6.98 (s, 1H), 6.99-7.02 (m, 1H), 7.17 (s, 1H), 7.44 (s, 1H),7.57 (d, J=5.0 Hz, 1H), 8.03 (d, J=10.0 Hz, 1H), 8.14 (s, 1H);

LC-MS (retention time: 1.92 min, method B), MS m/z 780 (M⁺+H).

Example 42 Preparation of Compound 42

Prepared following the procedures used in the preparation of Example 11.Compound 11, except that 6-ethoxy cinnamic acid was used in place of6-methoxy cinnamic acid as starting material for the P2 element.

¹H NMR (500 MHz, CD₃OD) δ ppm 0.98-1.09 (m, 15H), 1.24-1.31 (m, 10H),1.42-1.46 (m, 1H), 1.85-1.90 (m, 1H), 2.19-2.32 (m, 2H), 2.57-2.63 (m,1H), 2.91-2.97 (m, 1H), 4.03-4.09 (m, 1H), 4.17 (q, J=7.0 Hz, 2H), 4.42(d, J=11.3 Hz, 1H), 4.49-4.54 (m, 1H), 5.12 (d, J=17.4 Hz, 1H),5.72-5.78 (m, 1H), 5.83 (s, 1H), 7.07-7.10 (M, 1H), 7.15 (s, 1H), 7.22(d, J=5.8 Hz, 1H), 7.87 (d, J=5.8 Hz, 1H), 8.08 (d, J=8.8 Hz, 1H); MS:(M+H)⁺ 728.

Section C Example 45 Preparation of Compound 45

Step 1:

To a solution of 2-bromo-5-methoxybenzoic acid (1.68 g, 7.27 mmol) inDMF (50 mL) in a medium pressure flask (Chemglass) was added benzamidine(1.25 g, 8.00 mmol), K₂CO₃ (6.0 g, 43.6 mmol), and copper powder (336mg, 1.45 mmol). The reaction mixture was heated to 180° C. for 1 h.Copper and excess K₂CO₃ were removed by vacuum filtration and washedwith MeOH. The filtrate was concentrated and the resulting crude waspurified by flash column chromatography (SiO₂, 5% MeOH in DCM) to give alight green solid (1.55 g, 84% yield): ¹H NMR (DMSO-d₆) δ 3.84 (s, 3H),7.26 (d, J=7.8 Hz, 1H), 7.46 (br s, 5H), 7.57 (s, 1H), 8.38 (br s, 1H);MS m/z (MH⁺) 253.

Step 2:

To a 0° C. slurry of Boc-cis-Hydroxyproline-OMe (2.0 g, 8.15 mmol) andthe product from Example 45, Step 1 (2.26 g, 8.97 mmol) in THF (82 mL)was added Ph₃P and diisopropyl azocarboxylate (1.98 g, 8.97 mmol). Afterstirring at rt for 17 h, the reaction mixture was diluted with EtOAc(100 mL) and washed with H₂O (50 mL). The aqueous layer was separatedand back-extracted with EtOAc (2×50 mL). The combined organic layer waswashed with brine, dried over MgSO₄ and concentrated to give a viscousoil which was redissolved in minimal amount of EtOAc and hexanes wasadded to effect the precipitation of most of the Ph₃PO by-product. Ph₃POwas removed by vacuum filtration and the liquid filtrate wasconcentrated. The resulting viscous oil was purified by a flash columnchromatography (SiO₂, 4:1 hex:EtOAc) to give a white solid product (1.76g, 45% yield): ¹H NMR (60/40 rotomers, CDCl₃) δ 1.47 (s, 9H), 2.49-2.55(m, 1H), 2.73-2.83 (m, 1H), 3.80 (s, 1.8H), 3.81 (s, 1.2H), 3.96 (s,3H), 4.03-4.09 (m, 1H), 4.54 (t, J=8.0 Hz, 0.6H), 4.66 (t, J=7.8 Hz),4.96-5.06 (m, 1H), 5.97 (br s, 0.6H), 6.04 (br s, 0.4H), 7.33 (dd,J=6.1, 2.7 Hz, 1H), 7.46-7.51 (m, 4H), 7.91 (d, J=9.2 Hz, 1H), 8.49 (t,J=8.5 Hz, 2H); ¹³C NMR (rotomers, CDCl₃) δ 21.7, 22.0, 28.3, 28.4, 35.8,36.8, 52.3, 52.4, 52.6, 55.8, 55.9, 57.9, 58.3, 74.5, 74.9, 80.6, 101.2,101.3, 115.7, 125.8, 126.0, 128.1, 128.5, 129.7, 130.2, 137.9, 147.8,153.8, 157.7, 158.0, 158.0, 164.8, 173.1, 173.3; MS m/z (MH⁺) 480.

Step 3:

The product from Example 45, Step 2 (760.0 mg, 1.59 mmol) was dissolvedin 50% TFA in DCM and stirred at rt for 2 h. The solvent wasconcentrated and the resulting brown viscous oil was dried in vacuoovernight. The product was used directly for the next reaction.

Step 4:

To a solution of the brown viscous oil product from Example 45, Step 3(963 mg, 1.59 mmol) and DIPEA (1.23 g, 9.54 mmol) in DCM (11 mL) wereadded N—BOC L-tBuGly (440 mg, 1.90 mmole), HBTU (902 mg, 2.38 mmole) andHOBt (364 mg, 2.38 mmole). After stirring at rt for 14 h, the solventand excess DIPEA was concentrated and the resulting brown viscous oilwas purified by flash column (SiO₂, 4:1 hex:EtOAc) to give a white solid(0.922 mg, 98% yield for the two steps): ¹H NMR (CDCl₃/MeOD) δ 0.94 (s,9H), 1.15 (s, 9H), 2.38-2.42 (m, 1H), 2.60-2.73 (m, 1H), 3.61 (s, 3H),3.83 (s, 3H), 4.08-4.17 (m, 2H), 4.25 (d, J=11.5 Hz, 1H), 4.69 (t, J=8.0Hz, 1H), 5.99 (br s, 1H), 7.13 (s, 1H), 7.38 (s, 5H), 7.80 (d, J=9.0 Hz,1H), 8.32 (d, J=5.5 Hz, 1H); ¹³C NMR (CDCl₃/MeOD) δ 29.6, 31.4, 31.6,33.04, 38.2, 39.0, 55.8, 56.9, 59.2, 61.5, 62.1, 78.3, 83.1, 105.0,119.0, 129.4, 131.5, 131.9, 132.6, 133.8, 141.2, 151.0, 161.4, 161.6,168.2, 175.2, 175.7; MS m/z (MH⁺) 593.

Step 5:

To a solution of the product from Example 45, Step 4 (409 mg, 0.69 mmol)in THF (10 mL) was added 1N NaOH (2 mL). After stirring at rt for 19 h,the reaction was acidified with concentrated HCl to about pH 5 andextracted with DCM (3×50 mL). The combined organic layer was dried overMgSO₄ and concentrated to give a yellow solid product (370 mg, 92%yield) which was used directly in the next reaction after drying invacuo: ¹H NMR (CDCl₃) δ 1.05 (s, 9H), 1.25 (s, 9H), 2.76-2.83 (m, 2H),3.94 (s, 3H), 4.23-4.27 (m, 2H), 4.41 (d, J=11.6 Hz, 1H), 4.92 (t, J=7.6Hz, 1H), 5.20 (d, J=8.9 Hz, 1H), 6.08 (br s, 1H), 7.31 (s, 1H),7.46-7.50 (m, 5H), 7.93 (d, J=9.15 Hz, 1H), 8.51 (d, J=7.3 Hz, 2H); MSm/z (MH⁺) 579.

Step 6:

To a solution N-Boc-vinylcyclopropanecarboxylic acid (1R,2S/1S,2R 1:1mixture) (1.01 g, 4.46 mmol) in THF (20 mL) and DMSO (2 mL) was addedCDI (1.08 g, 6.69 mmol) and DMAP (817 mg, 6.69 mmol). After stirring at70° C. for 1 h, the reaction mixture was allowed to cool to rt and wastreated with isopropylsulfonamide (1.1 g, 8.92 mmol) and DBU (1.36 g,8.92 mmol). The reaction mixture was stirred at rt for 16 h and it wasconcentrated and purified by flash column chromatography (SiO₂, 5% MeOHin DCM) to give a brown viscous oil (1.4 g, 98% yield): ¹H NMR(Methanol-d₄) δ 1.25 (m, 1H), 1.33 (d, J=6.7 Hz, 3H), 1.36 (d, J=6.7 Hz,3H), 1.45 (s, 9H), 1.84 (dd, J=7.6, 5.2 Hz, 1H), 2.16 (d, J=7.6 Hz, 1H),3.58 (br s, 1H), 5.08 (d, J=11.6 Hz, 1H), 5.27 (d, J=15.6 Hz, 1H),5.58-5.66 (m, 1H); MS m/z (MH⁺) 332.

Step 7:

The product from Example 45, Step 6 (113 mg, 0.34 mmol) was treated witha 50% solution of trifluoroacetic acid in DCM (10 mL) and stirred at rtfor 1.4 h. Solvent and excess trifluoroacetic acid were removed invacuo. The resulting brown viscous oil was dried in vacuo (1.3 g, 99%yield) and used without further purification: ¹H NMR (DMSO-d₆) δ 1.24(d, J=6.7 Hz, 3H), 1.26 (d, J=6.7 Hz, 3H), 1.54 (dd, J=9.6, 6.6 Hz, 1H),1.99 (t, J=6.9 Hz, 1H), 2.24 (d, J=8.5 Hz, 1H), 3.58-3.63 (m, 1H), 5.18(d, J=10.4 Hz, 1H), 5.33 (d, J=17.1 Hz, 1H), 5.61-5.69 (m, 1H), 8.83 (brs, 3H); ¹³C NMR (DMSO-d₆) δ 15.2, 15.9, 16.5, 29.9, 41.6, 52.1, 116.0,118.9, 132.0, 158.2, 167.3; MS m/z (MH⁺) 233.

Step 8:

To a mixture of the product from Example 45, Step 5 (117 mg, 0.338 mmol)and DIPEA (174 mg, 1.35 mmol) in DCM (5 mL) was added HBTU (128 mg,0.338 mmole), HOBt (52 mg, 0.338 mmole) and the product from Example 45,Step 7 (130 mg, 0.225 mmol) After stirring at rt for 16 h, the mixturewas concentrated and the resulting brown viscous oil was purified byflash column chromatography (SiO₂, 1:3 hex:EtOAc then 95:5 DCM:MeOH) togive an off white solid product (150 mg, 84% yield) The final product,Compound 45, is a mixture of isomers; the variation occurring at the P1vinylcyclopropyl portion of the molecule (1R,2S/1S,2R 1:1 mixture): ¹HNMR (Methanol-d₄) δ 0.92 (br s, 2H), 1.03 (s, 9H), 1.17 (s, 9H),1.27-1.38 (m, 9H), 1.42-1.46 (m, 1H), 1.83 (dd, J=8.1, 5.3 Hz, 0.4H),1.90 (dd, J=7.9, 5.5 Hz, 0.6H), 2.24-2.31 (m, 1H), 2.37-2.45 (m, 1H),2.67-2.75 (m, 1H), 3.73-3.79 (m, 1H), 3.90 (s, 3H), 4.21 (dd, J=9.3, 6.0Hz, 2H), 4.48 (d, J=11.3 Hz, 1H), 4.61 (q, J=8.9 Hz, 1H), 5.14 (t, J=9.0Hz, 1H), 5.33 (t, J=17.9 Hz, 1H), 5.70-5.76 (m, 1H), 6.06 (d, J=11.9 Hz,1H), 6.61 (d, J=8.9 Hz, 1H), 7.34 (d, J=2.8 Hz, 1H), 7.49 (br s, 5H),7.87 (d, J=8.9 Hz, 1H), 8.46 (d, J=4.3 Hz, 2H); ¹³C NMR (Methanol-d₄) δ15.7, 16.1, 16.5, 16.8, 23.9, 27.1, 28.6, 35.8, 36.0, 36.2, 36.3, 36.4,42.6, 42.8, 54.7, 54.8, 55.5, 56.4, 61.1, 61.2, 80.5, 102.9, 117.0,118.8, 118.9, 126.8, 129.4, 129.6, 130.2, 131.5, 134.4, 139.2, 148.8,158.0, 159.3, 159.8, 166.3, 171.1, 175.1, 184.3; MS m/z (MH⁺) 793.

Example 46 Preparation of Compound 46

Compound 46 was prepared by following Steps 1 through 5 and Step 8 ofExample 45 except that the following modifications were made:

Step 1:

Modifications: 2-bromo-4,5-dimethoxybenzoic acid andcyclopropylcarbamidine hydrochloride were utilized as startingmaterials.

Product:

Data: ¹H NMR (DMSO-d₆) δ 0.97-1.01 (m, 2H), 1.03-1.06 (m, 2H), 1.90-1.94(m, 1H), 3.84 (s, 3H), 3.87 (s, 3H), 6.93 (s, 1H), 7.37 (s, 3H), 12.28(s, 1H); ¹³C NMR (DMSO-d₆) δ 9.03, 13.17, 55.47, 55.73, 104.81, 107.27,113.26, 145.16, 147.48, 154.44, 157.21, 160.89; MS m/z (MH⁺) 247.

Step 2:

Modifications: The product from Example 46, Step 1 was used as startingmaterial in place of the product from Example 45, Step 1.

Product:

Data: ¹H NMR (CDCl₃) δ 1.00-1.04 (m, 2H), 1.07-1.11 (m, 2H), 1.43 (s,5.4H), 1.46 (s, 3.6H), 2.17-2.21 (m, 1H), 2.37-2.43 (m, 1H), 2.62-2.69(m, 1H), 3.75 (s, 1.8H), 3.78 (s, 1.2H), 3.92 (d, J=2.8 Hz, 1H), 4.00(s, 3.6H), 4.01 (s, 2.4H), 4.48 (t, J=8.0 Hz, 0.6H), 4.59 (t, J=7.6 Hz,0.4H), 5.7 (br s, 0.6H), 5.74 (br s, 0.4H), 7.18 (s, 1H), 7.20 (s, 1H);¹³C NMR (CDCl₃) δ 9.6, 9.7, 18.1, 28.3, 28.4, 35.8, 36.7, 52.2, 52.4,56.3, 57.8, 58.2, 74.0, 74.5, 80.5, 80.6, 101.0, 101.1, 106.3, 108.6,148.8, 149.1, 153.8, 155.4, 164.4, 165.9, 172.9, 173.2; LC-MS m/z (MH⁺)474.

Steps 3 and 4:

The product from Example 46, Step 2 was used as starting material inplace of the product from Example 45, Step 2.

Product:

Data: ¹H NMR (Methanol-d₄) δ 1.04 (s, 9H), 1.08-1.21 (m, 4H), 1.14 (s,9H), 2.17-2.21 (m, 1H), 2.39-2.41 (m, 1H), 2.74-2.77 (m, 1H), 3.77 (s,3H), 3.92 (s, 3H), 3.98 (m, 3H), 4.09 (dd, J=11.4, 3.8 Hz, 1H), 4.17 (d,J=8.9 Hz, 1H), 4.42 (d, J=11.3 Hz, 1H), 4.76 (t, J=8.2 Hz, 1H), 5.81 (brs, 1H), 6.43 (d, J=8.6 Hz, 1H), 7.14 (d, J=6.1 Hz, 1H), 7.27 (d, J=5.8Hz, 1H); ¹³C NMR (Methanol-d₄) δ 10.0, 10.3, 18.6, 26.9, 28.5, 28.8,35.8, 36.1, 38.9, 52.8, 54.9, 56.7, 59.6, 60.5, 76.6, 80.4, 102.7,106.2, 109.9, 149.8, 150.7, 157.6, 166.0, 167.3, 173.5, 173.6; MS m/z(MH⁺) 587.

Step 5:

The product from Example 46, Step 4 was used as starting material inplace of the product from Example 45, Step 4.

Product:

Data: ¹H NMR (Methanol-d₄) δ 1.03 (s, 9H), 1.13 (s, 9H), 1.20-1.23 (m,4H), 2.15-2.19 (m, 1H), 2.40-2.45 (m, 1H), 2.70-2.76 (m, 1H), 3.90 (s,3H), 3.96 (s, 3H), 4.08 (dd, J=11.4, 3.8 Hz, 1H), 4.17 (d, J=5.8 Hz,1H), 4.37 (d, J=11.3 Hz, 1H), 4.71 (t, J=8.1 Hz, 1H), 5.77 (br s, 1H),7.09 (s, 1H), 7.20 (s, 1H); ¹³C NMR (Methanol-d₄) δ 10.2, 10.5, 18.6,26.9, 28.5, 28.8, 36.0, 36.3, 54.9, 56.8, 59.7, 60.4, 76.8, 80.4, 102.6,105.9, 109.9, 126.9, 127.9, 149.3, 150.8, 157.65, 157.8, 166.1, 167.3,173.3, 175.1; MS m/z (MH⁺) 573.

Step 8:

The product from Example 46, Step 5 was used as starting material inplace of the product from Example 45, Step 5. The final product,Compound 46, is a mixture of isomers; the variation occurring at the P1vinylcyclopropyl portion of the molecule (1R,2S/1S,2R 1:1 mixture).

Product:

Data: ¹H NMR (Methanol-d₄) δ 1.03 (s, 9H), 1.05-1.09 (m, 4H), 1.16 (s,4.5H), 1.17 (s, 4.5H), 1.19-1.22 (m, 1H), 1.31 (d, J=6.7 Hz, 2H),1.33-1.38 (m, 7H), 1.18-1.89 (m, 1H), 2.15-2.20 (m, 2H), 2.35-2.44 (m,1H), 3.23 (q, J=7.4 Hz, 1H), 3.70-3.75 (m, 1H), 3.91 (s, 3H), 3.98 (s,3H), 4.08-4.13 (m, 2H), 4.16 (dd, J=8.9, 3.1 Hz, 1H), 4.38 (t, J=13.1Hz, 1H), 4.58-4.62 (m, 1H), 4.06 (m, 1H), 5.29 (t, J=15.2 Hz, 1H), 5.83(br s, 1H), 7.15 (s, 1H), 7.27 (d, J=4.3 Hz, 1H); MS m/z (MH⁺) 787.

Example 47 Preparation of Compound 47

Compound 47 was prepared by following analogous steps used to procureCompound 45 of Example 45. ortho-bromobenzoic acid used as startingmaterial as was cyclopropanesulfonic acid(1R-amino-2S-vinyl-cyclopropanecarbonyl)-amide hydrochloride salt.Compound 47: MH+=761

Example 48 Preparation of Compound 48

Compound 48 was prepared by following Steps 1 through 5 and Step 8 ofExample 45 except that the following modifications were made:

Step 1:

Modifications: Acetamidine hydrochloride and 2-bromo-5-methoxybenzoicacid were utilized as starting materials.

Product:

Data: ¹H NMR (DMSO) δ 2.31 (s, 3H), 3.85 (s, 3H), 7.36 (d, J=6.2 Hz,1H), 7.37 (s, 1H), 7.51 (d, J=7.8 Hz, 1H), 12.15 (s, 1H); ¹³C NMR (DMSO)δ 21.11, 55.41, 105.57, 121.22, 123.59, 128.12, 143.34, 151.68, 157.00,161.45; LC-MS m/e (MH⁺) 191.

Step 2:

Modifications: The product from Example 48, Step 1 was used as startingmaterial in place of the product from Example 45, Step 1.

Product:

Data: ¹H NMR (CDCl₃) δ 1.43 (s, 5.4H), 1.45 (s, 3.6H), 2.38-2.45 (m,1H), 2.62-2.71 (m, 1H), 2.66 (s, 1.8H), 2.68 (s, 1.2H), 3.77 (1.8H),3.79 (s, 1.2H), 3.92 (s, 3H), 3.93-3.98 (m, 2H), 4.49 (t, J=8.0 Hz,0.6H), 4.61 (t, J=7.8 Hz, 0.4H), 5.82 (t, J=2.1 Hz, 0.6H), 5.89 (t,J=2.3 Hz, 0.4H), 7.26 (dd, J=4.7, 3.2 Hz, 1H), 7.42 (dd, J=6.3, 2.8 Hz,1H), 7.75 (d, J=9.15 Hz, 1H); ¹³C NMR (CDCl₃) δ 26.1, 28.3, 28.4, 35.8,36.7, 52.2, 52.2, 52.4, 52.5, 55.7, 55.8, 57.9, 58.2, 74.1, 74.7, 80.6,101.0, 101.2, 114.9, 125.6, 125.9, 128.6, 147.3, 153.8, 154.5, 157.6,157.6, 161.2, 164.6, 173.0, 173.3; LC-MS m/e (MH⁺) 418.

Steps 3 and 4:

Modifications: The product from Example 48, Step 2 was used as startingmaterial in place of the product from Example 45, Step 2.

Product:

Data: ¹HNMR (MeOD) δ 1.03 (s, 9H), 1.07 (s, 9H), 2.38-2.42 (m, 1H), 2.68(s, 3H), 2.80 (q, J=7.8 Hz, 1H), 3.76 (s, 3H), 3.89 (s, 3H), 4.07 (dd,J=11.9, 3.4 Hz, 1H), 4.13 (br s, 1H), 4.55 (d, J=12.2 Hz, 1H), 4.78 (t,J=8.7 Hz, 1H), 5.93 (s, 1H), 7.37 (d, J=2.75 Hz, 1H), 7.48-7.51 (m, 2H),7.70 (d, J=5.7 Hz, 1H); ¹³C NMR (MeOD) δ 25.6, 26.9, 28.4, 28.8, 35.9,52.8, 55.0, 56.4, 59.7, 60.6, 77.2, 80.4, 102.9, 111.6, 116.5, 127.0,128.4, 147.5, 162.7, 166.4, 173.6; LC-MS m/e (MH⁺) 531.

Step 5:

Modifications: The product from Example 48, Step 4 was used as startingmaterial in place of the product from Example 45, Step 4.

Product:

Data: ¹H NMR (MeOD) δ 1.03 (s, 9H), 1.08 (s, 9H), 2.41-2.46 (m, 1H),2.68 (s, 3H), 2.81 (q, J=8.1 Hz, 1H), 3.89 (s, 3H), 4.07 (dd, J=11.8,3.2 Hz, 1H), 4.18 (d, J=5.5 Hz, 1H), 4.52 (d, J=11.9 Hz, 1H), 4.74 (t,J=8.7 Hz, 1H), 5.93 (br s, 1H), 7.37 (d, J=2.81 Hz, 1H), 7.49 (dd,J=9.2, 2.4 Hz, 1H), 7.71 (d, J=9.2 Hz, 1H); ¹³C NMR (MeOD) δ 25.7, 26.9,28.5, 36.1, 55.0, 56.4, 59.7, 60.5, 77.1, 80.4, 103.0, 116.5, 127.0,128.5, 147.7, 157.8, 159.6, 162.7, 166.4, 173.5, 174.9; LC-MS m/e (MH⁺)517.

Example 48 Preparation of Compound 48

Step 8:

To a solution of the product from Example 48, Step 5 (45.8 mg, 0.089mmol), cyclopropanesulfonic acid(1R-amino-2S-vinyl-cyclopropanecarbonyl)-amide hydrochloride salt (21.0mg, 0.089 mmol) and DIEA (34.5 mg, 0.267 mmol) in DCM (1 mL) was addedHATU (44.0 mg, 0.116 mmol). After stirring at rt overnight, the reactionmixture was washed with 5% aqueous NaHCO₃ (1 mL). The aqueous layer wasextracted was 2×2 mL DCM. The organic layer was washed with 5% aqueouscitric acid (1 mL), brine, dried over MgSO₄, concentrated and purifiedby reversed prep-HPLC. This purification step resulted in the loss ofthe N—BOC protecting group at the P3 tert-leucine portion of themolecule: ¹H NMR (MeOD) δ 1.07-1.12 (m, 2H) 1.14 (s, 2H) 1.14-1.16 (m,2H) 1.17 (s, 9H) 1.20-1.30 (m, 3H) 1.45 (dd, J=9.46, 5.49 Hz, 1H) 1.56(s, 1H) 1.92 (dd, J=8.20, 5.60 Hz, 1H) 2.25-2.31 (m, 1H) 2.39-2.45 (m,1H) 2.73 (m, 1H) 2.76 (s, 3H) 2.93-2.97 (m, 1H) 3.94 (s, 1H) 3.96 (s,3H) 4.07 (s, 1H) 4.21 (d, J=3.97 Hz, 0.4H) 4.23 (d, J=3.97 Hz, 0.6H)4.31 (m, 1H) 4.73 (dd, J=10.38, 7.02 Hz, 1H) 5.15 (dd, J=10.38, 1.52 Hz,1H) 5.32 (dd, J=17.1, 1.52 Hz, 1H), 5.71-5.78 (m, 1H) 6.11 (t, J=3.51Hz, 1H) 7.46 (d, J=2.75 Hz, 1H) 7.67 (d, J=3.06 Hz, 0.4H) 7.69 (d,J=3.05 Hz, 0.6H) 7.82 (s, 0.6H) 7.84 (s, 0.4H).

Example 49 Preparation of Compound 49

Compound 49 was prepared by the same method as described for thepreparation of Compound 48, except the product from Example 46, Step 5and cyclopropanesulfonic acid(1R-amino-2S-vinyl-cyclopropanecarbonyl)-amide hydrochloride salt wereused as starting material. The preparative HPLC purification stepresulted in the loss of the N—BOC protecting group at the P3tert-leucine portion of the molecule: ¹H NMR (MeOD) δ 1.09 (m, 2H) 1.14(d, J=3.97 Hz, 2H) 1.17 (s, 9H) 1.25 (m, 3H) 1.37 (m, 3H) 1.44 (dd,J=9.31, 5.65 Hz, 2H) 1.57 (s, 1H) 1.92 (dd, J=8.09, 5.65 Hz, 1H) 2.28(dd, J=17.70, 8.55 Hz, 1H) 2.32 (m, 1H) 2.68 (dd, J=14.19, 7.78 Hz, 1H)2.95 (m, 1H) 3.98 (s, 3H) 4.06 (s, 3H) 4.08 (s, 1H) 4.22 (d, J=2.75 Hz,1H) 4.70 (dd, J=9.77, 7.32 Hz, 1H) 5.15 (dd, J=10.38, 1.53 Hz, 1H) 5.32(dd, J=17.40, 1.22 Hz, 1H) 5.74 (m, 1H) 6.04 (m, 1H) 7.24 (s, 1H) 7.37(s, 1H)

Example 50 Preparation of Compound 50

Compound 50 was prepared by the same method as described for thepreparation of Compound 48, except the product from Example 45, Step 5and cyclopropanesulfonic acid(1R-amino-2S-vinyl-cyclopropanecarbonyl)-amide hydrochloride salt wereused as starting material. The preparative HPLC purification stepresulted in the loss of the N—BOC protecting group at the P3tert-leucine portion of the molecule: ¹H NMR (MeOD) δ 1.10 (m, 2H) 1.14(s, 1H) 1.15 (d, J=3.36 Hz, 1H) 1.17 (d, J=3.05 Hz, 9H) 1.22 (m, 1H)1.27 (m, 2H) 1.46 (dd, J=9.46, 5.49 Hz, 1H) 1.56 (s, 1H) 1.93 (dd,J=8.24, 5.49 Hz, 1H) 2.29 (q, J=8.55 Hz, 1H) 2.48 (m, 1H) 2.78 (dd,J=13.89, 8.09 Hz, 1H) 2.97 (m, 1H) 3.96 (s, 2H) 4.07 (s, 1H) 4.32 (d,J=2.14 Hz, 2H) 4.76 (d, J=7.02 Hz, 1H) 4.78 (m, 1H) 4.86 (d, J=3.05 Hz,1H) 5.32 (dd, J=17.09, 1.22 Hz, 1H) 5.75 (m, 1H) 6.24 (d, J=2.44 Hz, 1H)7.45 (d, J=2.75 Hz, 1H) 7.52 (m, 3H) 7.61 (dd, J=9.16, 2.75 Hz, 1H) 7.96(d, J=9.16 Hz, 1H).

Example 51 Preparation of Compound 51

Compound 51 was prepared by following Steps 1 through 5 and Step 8 ofExample 45 except that the following modifications were made:

Step 1:

Modifications: 2-bromo-4,5-dimethoxybenzoic acid and trifluoroamidinewere utilized as starting materials.

Product:

Data: ¹H NMR (DMSO) δ 3.92 (s, 3H), 3.94 (s, 3H), 7.33 (s, 1H), 7.50 (s,1H), 13.40 (br s, 1H); ¹³C NMR (DMSO) δ 55.8, 56.1, 104.9, 108.7, 150.2,155.0; LC-MS m/e (MH⁺) 275.

Step 2:

Modifications: The product from Example 51, Step 1 was used as startingmaterial in place of the product from Example 45, Step 1.

Product:

Data: ¹H NMR (CDCl₃) δ 1.42 (s, 3.6H), 1.44 (s, 5.4H), 2.42-2.49 (m,1H), 2.67-2.73 (m, 1H), 3.37 (s, 1.2H), 3.78 (s, 1.8H), 3.97 (t, J=6.5Hz, 1H), 4.02 (s, 2.4H), 4.04 (s, 3.6H), 4.48 (t, J=7.9 Hz, 0.6H), 4.60(t, J=7.7 Hz, 0.4H), 5.86 (br s, 0.6H), 5.90 (br s, 0.4H), 7.27-7.29 (m,1H), 7.38-7.44 (m, 1H); ¹³C NMR (CDCl₃) δ 8.2, 28.3, 35.7, 36.7, 52.1,52.2, 52.4, 56.5, 57.8, 58.2, 75.5, 76.0, 80.7, 100.8, 107.6, 111.0,119.7, 148.2, 150.2, 151.4, 153.8, 154.5, 156.4, 165.1, 172.7, 173.0;LC-MS m/e (MH⁺) 502.

Steps 3 and 4:

Modifications: The product from Example 51, Step 2 was used as startingmaterial in place of the product from Example 45, Step 2.

Product:

Data: ¹H NMR (MeOD) δ 1.03 (s, 9H), 1.08 (s, 9H), 2.41-2.45 (m, 1H),2.80-2.84 (m, 1H), 3.76 (s, 3H), 3.96 (s, 3H), 4.00 (s, 3H), 4.10-4.14(m, 2H), 4.52 (d, J=11.6 Hz, 1H), 4.80 (t, J=8.7 Hz, 1H), 5.92 (br s,1H), 7.35 (br s, 2H); ¹³C NMR (MeOD) 26.9, 28.4, 28.8, 35.7, 36.0, 52.8,54.8, 56.9, 59.6, 60.7, 77.9, 80.3, 102.2, 107.9, 112.4, 120.3, 149.3,153.2, 157.8, 158.3, 173.5; LC-MS m/e (MH⁺) 615.

Step 5:

Modifications: The product from Example 51, Step 4 was used as startingmaterial in place of the product from Example 45, Step 4.

Product:

Data: ¹H NMR (MeOD) δ 1.03 (s, 9H), 1.09 (s, 9H), 2.44-2.49 (m, 1H),2.80-2.84 (m, 1H), 3.97 (s, 3H), 4.01 (s, 3H), 4.10-4.24 (m, 3H), 4.50(d, J=11.9 Hz, 1H), 4.76 (t, J=7.9 Hz, 1H), 5.93 (br s, 1H), 7.36 (br s,2H); ¹³C NMR (MeOD) δ 26.9, 28.4, 28.8, 36.0, 36.1, 54.8, 56.9, 57.0,60.6, 77.9, 80.3, 102.3, 108.0, 112.5, 120.3, 149.3, 151.3, 153.2,158.2, 158.3, 166.7, 173.5; LC-MS m/e (MH⁺) 601.

Step 8:

Modifications: The product from Example 51, Step 5 was used as startingmaterial in place of the product from Example 45, Step 5. The finalproduct, Compound 51, is a mixture of isomers; the variation occurringat the P1 vinylcyclopropyl portion of the molecule (1R,2S/1S,2R 1:1mixture).

Product:

Data: ¹H NMR (DMSO) δ 0.23 (s, 4.5H), 0.23 (s, 4.5H), 0.35 (s, 4.5H),0.36 (s, 4.5H), 0.45-0.59 (m, 8H), 0.63-0.66 (m, 1H), 1.04 (dd, J=8.2,5.2 Hz, 1H), 1.10 (dd, J=8.2, 5.5 Hz, 1H), 1.47-1.53 (m, 1H), 1.58-1.61(m, 1H), 1.87-1.90 (m, 1H), 2.95-3.01 (m, 1H), 3.17 (s, 1.5H), 3.18 (s,1.5H), 3.22 (s, 3H), 3.37 (br s, 2H), 3.68 (q, J=5.9 Hz, 1H), 3.82 (q,J=8.6 Hz, 1H), 4.33-4.37 (m, 1H), 4.54 (t, J=16.5 Hz, 1H), 4.93 (q,J=8.9 Hz, 1H), 5.17 (d, J=15.9 Hz, 1H), 6.53 (s, 1H), 6.58 (s, 1H); ¹³CNMR (DMSO) δ 12.8, 13.2, 13.7, 13.9, 19.5, 20.6, 21.1, 24.3, 25.6, 32.9,33.1, 33.4, 33.6, 36.1, 39.7, 39.9, 51.8, 51.9, 52.4, 54.0, 54.2, 57.7,57.7, 58.1, 58.3, 75.1, 75.3, 77.5, 84.1, 99.2, 105.2, 107.9, 109.5,116.0, 116.1, 118.7, 123.9, 127.4, 131.5, 146.5, 148.6, 150.3, 155.1,155.5, 163.7, 164.7, 168.2, 168.3, 170.7, 172.2; LC-MS m/e (MH⁺) 815.

Example 52 Preparation of Compound 52

Compound 52 was prepared by the same method as described for thepreparation of Compound 48, except the product from Example 51, Step 5and cyclopropanesulfonic acid(1R-amino-2S-vinyl-cyclopropanecarbonyl)-amide hydrochloride salt wereused as starting material. The preparative HPLC purification stepresulted in the loss of the N—BOC protecting group at the P3tert-leucine portion of the molecule: ¹H NMR (MeOD) δ 1.11 (m, 3H) 1.17(s, 9H) 1.25 (m, 3H) 1.46 (dd, J=9.46, 5.49 Hz, 1H) 1.92 (dd, J=8.24,5.49 Hz, 1H) 2.28 (q, J=8.95 Hz, 1H) 2.42 (m, 1H) 2.72 (dd, J=14.19,7.17 Hz, 1H) 2.96 (m, 1H) 4.01 (s, 3H) 4.04 (m, 5H) 4.24 (m, 2H) 4.73(dd, J=10.22, 7.17 Hz, 1H) 5.15 (dd, J=10.53, 1.37 Hz, 1H) 5.32 (d,J=17.09 Hz, 1H) 5.75 (m, 1H) 6.07 (s, 1H) 7.41 (s, 1H) 7.47 (s, 1H).

Example 53 Preparation of Compound 53

Step 1:

To a solution of (1R,2S/1S,2R 1:1 mixture) cyclopropanesulfonic acid(1-amino-2-vinyl-cyclopropanecarbonyl)-amide trifluoroacetic acid salt(626 mg, 1.82 mmol) in DCM (17 mL) was added and DIEA (555 mg, 4.29mmol) in DCM (17 mL), HATU (754 mg, 1.98 mmole), and(2S,4R)Fmoc-4-amino-1-boc-pyrrolidine-2-carboxylic acid (747 mg, 1.65mmol). After stirring at rt for 24 h, the mixture was washed with 1N HCl(10 mL), 5% aqueous NaHCO₃ (4 mL). Each aqueous layer was extracted withDCM (25 mL). The combined DCM was dried over MgSO₄ and concentrated. Theresulting brown viscous oil was purified by flash column chromatography(SiO₂, 95:5 DCM:MeOH) to give a yellow solid 822 mg, 75% yield):

¹H NMR (DMSO-d₆) δ 1.04-1.09 (m, 3H), 1.15-1.27 (m, 4H), 1.38-1.44 (m,7H), 1.47 (s, 9H), 1.84 (dd, J=8.2, 5.2 Hz, 1H), 2.01-2.30 (m, 4H),2.90-2.98 (m, 1H), 3.64-3.71 (m, 1H), 4.16-4.22 (m, 4H), 4.39 (bs, 2H),5.13 (dd, J=10.7, 0.9 Hz, 1H), 3.31 (d, J=17.1 Hz, 1H), 5.72-5.79 (m,1H), 7.31 (t, J=7.3 Hz, 3H), 7.38 (t, J=7.5 Hz, 3H), 7.64 (d, J=7.02 Hz,3H), 7.79 (d, J=7.63 Hz, 3H); LC-MS m/e (Na⁺ MH⁺) 687.

Step 2:

The product from Example 53, Step 1 (500 mg, 0.752 mmol) was treatedwith 50% TFA in DCM (10 mL). After stirring at rt for 0.5 h, theresulting brown reaction mixture was concentrated to give a brown solid(489 mg, 84% yield): ¹H NMR (DMSO-d₆) δ 1.03-1.19 (m, 4H), 1.24-1.26 (m,1H), 1.35 (dd, J=9.5, 5.5 Hz, 1H), 1.91-1.96 (m, 1H), 2.22-2.30 (m, 1H),2.40 (bs, 1H), 2.93-2.98 (m, 1H), 3.60 (bs, 1H), 4.21 (t, J=5.6 Hz, 2H),4.47 (bs, 3H), 5.17 (d, J=9.2 Hz, 1H), 5.32 (d, J=17.1 Hz, 1H),5.64-5.67 (m, 1H), 7.31 (t, J=7.3 Hz, 3H), 7.39 (t, J=7.5 Hz, 3H), 7.63(d, J=7.3 Hz, 2H), 7.80 (d, J=7.3 Hz, 2H); (LC-MS m/e (MH⁺) 565.

Step 3:

To a solution of the product from Example 53, Step 2 (260 mg, 0.383mmol) in DCM (4 mL) was added DIPEA (218 mg, 1.69 mmol), HATU (165 mg,0.422 mmol), and N—BOC L-tBuGly (100 mg, 0.422 mmol). After stirring atrt for 16 h, the reaction mixture was diluted with H₂O (3 mL), acidifiedwith 1N HCl to pH=1. The aqueous layer was extracted with DCM (2×15 mL).The combined organic layer was washed with 5% NaHCO₃ (3 mL), brine (5mL), dried over MgSO₄ and concentrated. The resulting brown viscous oilwas purified by flashed column chromatography (SiO₂, 95:5 DCM:MeOH) togive a brown foamy solid (281 mg, 94% yield): ¹H NMR (DMSO-d₆) δ0.96-1.08 (m, 4H), 1.05 (s, 9H), 1.15-1.26 (m, 2H), 1.35-1.38 (m, 5H),1.42 (s, 9H), 1.85 (dd, J=9.5, 5.5 Hz, 1H), 2.07 (bs, 1H), 2.22 (q,J=8.7 Hz, 1H), 2.92-2.95 (m, 1H), 3.90 (bs, 1H), 4.20 (d, J=6.4 Hz, 3H),4.29-4.39 (m, 5H), 5.13 (d, J=10.7, 1H), 5.31 (dd, J=18.0, 5.8 Hz, 1H),5.70-5.77 (m, 1H), 7.30 (t, J=7.3 Hz, 3H), 7.39 (t, J=7.3 Hz, 4H), 7.63(dd, J=6.7, 2.8 Hz, 3H), 8.80 (d, J=7.63 Hz, 3H); LC-MS m/z (MH⁺) 678.

Step 4:

The product from Example 53, Step 3 was treated with 10% piperidine inDMF (3.3 mL). After stirring at rt for 14 hr, solvent was removed andthe resulting brown viscous oil was purified by flash columnchromatography (SiO₂, 95:5 DCM:MeOH) to isolate the pure highest Rf1R,2S P1 diastereomer as a pale yellow solid (31 mg). The other isomerwas isolated in a mixture and was not used: LC-MS m/z (MH⁺) 556.

Steps 5 and 6:

To a solution of the product from Example 53, Step 4 in DMF (2 mL) wasadded polyvinylpyridine (13 mg) and Fmoc-isothiocyanate. After stirringat rt for 14 h, the reaction mixture was treated with piperidine (172mg, 2.02 mmol). The reaction was stirred at rt for an additional 6 hafter which it was concentrated and dried under vacuo overnight. Thecrude residue was re-dissolved in DMF (2 mL) and treated with2-bromoacetophenone and stirred at rt for another 14 h. The reactionmixture was concentrated and the resulting residue was purified by flashcolumn chromatography (SiO₂, 95:5 DCM:MeOH) to give Compound 53 as alight yellow solid (21.9 mg, 50% yield): ¹H NMR (DMSO-d₆) δ 0.87-0.92(m, 1H), 1.05 (bs, 13H), 1.16-1.25 (m, 4H), 1.34-1.38 (m, 2H), 1.42 (s,9H), 1.87 (t, J=6.6 Hz, 1H), 2.22-2.25 (m, 2H), 2.48 (t, J=10.7 Hz, 1H),2.93 (bs, 1H), 3.04 (q, J=7.3 Hz, 1H) 3.30-3.31 (m, 2H), 3.43-3.49 (m,1H), 4.01 (d, J=10.4 Hz, 1H), 4.07-4.12 (m, 1H), 4.27 (t, J=9.5 Hz, 1H),4.44 (t, J=7.0 Hz, 1H), 4.58 (bs, 1H), 5.11 (d, J=10.1 Hz, 1H), 5.30(dd, J=16.8, 9.6 Hz, 1H), 5.73-5.78 (m, 1H), 6.69 (d, J=8.2 Hz, 1H),6.86 (s, 1H), 7.25 (t, J=7.3 Hz, 1H), 7.35 (t, J=7.63 Hz, 2H), 7.82 (d,J=8.2 Hz, 2H); LC-MS m/z (MH⁺) 715.

Reversed Phase Prep-HPLC Conditions for Compound 55 Through 155 asIndicated:

Waters Xterra Prep MS C18 column, 5 mm (this means 5 micron particlesize), 30 mm×100 mm

Solvent A: 10% MeOH, 90% H₂O, 10 mM NH₄OAc

Solvent B: 90% MeOH, 10% H₂O, 10 mM NH₄OAc

50 mL/min flow rate

Gradient: 0% B to 100% B for 10 min, hold at 100% B for 4 min

Example 55 Preparation of Compound 55

Step 1:

To a solution of Compound 11 (1.5 g, 2.10 mmol) in DCE (25 mL) was addedTFA (25 mL). After stirring at rt for 15 min, the reaction mixture wasconcentrated. The resulting red viscous oil was re-dissolved in DCE (50mL) and reconcentrated. It was then redissolved in DCM (15 mL) andtreated with a solution of 1N HCl in Et₂O (25 mL). The resultingsuspension was chilled at 0° C., vacuum filtrated, washed with Et₂O anddried in vacuum oven to give product of step 1 as a white solid (1.4 g,97% yield): ¹HNMR (CD₃OD, 500 MHz) δ 1.07-11.2 (m, 3H) 1.14 (t, J=4.12Hz, 1H) 1.17 (s, 9H) 1.22 (dd, J=10.53, 4.43 Hz, 1H) 1.21-1.27 (m, 2H)1.42 (dd, J=9.61, 5.34 Hz, 1H) 1.91 (dd, J=7.93, 5.49 Hz, 1H) 2.27 (q,J=8.85 Hz, 1H) 2.32-2.38 (m, 1H) 2.70 (dd, J=13.43, 6.71 Hz, 1H),2.93-2.98 (m, 1H) 3.96 (s, 3H) 4.09 (s, 1H) 4.14 (dd, J=12.21, 3.66 Hz,1H) 4.32-4.35 (m, 1H) 4.69 (dd, J=10.53, 6.87 Hz, 1H) 5.14 (dd, J=10.38,1.53 Hz, 1H) 5.31 (dd, J=17.40, 1.22 Hz, 1H) 5.70-5.77 (m, 1H) 5.90 (t,J=3.51 Hz, 1H) 7.24-7.27 (m, 1H) 7.29 (d, J=4.27 Hz, 1H) 7.39 (t, J=4.88Hz, 1H) 7.90 (d, J=6.10 Hz, 1H) 8.19 (m, 1H) 9.22 (s, 1H).

Step 2:

To a solution mixture of product from step 1 of Example 55 (70.0 mg,0.108 mmol) and DIEA (41.8 mg, 0.323 mmol) in DCM (2 mL) was addedacetic anhydride (33.0 mg, 0.323 mmol). After stirring at rt for 14 h,solvent was removed and product was purified by reversed phase prep-HPLCto give Compound 55 (39.1 mg, 14% yield): ¹HNMR (CD₃OD, 500 MHz) δ1.00-1.03 (m, 1H), 1.06 (s, 9H), 1.07-1.10 (m, 1H), 1.21-1.28 (m, 2H),1.43 (dd, J=9.46, 5.19 Hz, 1H), 1.88 (dd, J=8.55, 5.49 Hz, 1H), 2.23 (q,J=8.85 Hz, 1H), 2.27-2.32 (m, 1H), 2.59 (dd, J=13.73, 7.02 Hz, 1H),2.92-2.97 (m, 1H), 3.93 (s, 3H), 4.12 (dd, J=11.90, 3.97 Hz, 1H), 4.35(d, J=11.60 Hz, 1H), 4.51 (dd, J=10.38, 7.02 Hz, 1H), 4.61 (dd, J=5.80,3.05 Hz, 1H), 4.80 (d, J=4.27 Hz, 1H), 4.88 (d, J=3.96 Hz, 1H), 5.12(dd, J=10.38, 1.83 Hz, 1H), 5.29 (dd, J=17.24, 1.37 Hz, 1H), 5.73-5.78(m, 1H), 5.84 (t, J=3.66 Hz, 1H), 7.15 (dd, J=8.85, 2.44 Hz, 1H), 7.19(d, J=2.44 Hz, 1H), 7.25 (d, J=6.10 Hz, 1H), 7.88 (d, J=6.10 Hz, 1H),8.06 (d, J=9.16 Hz, 1H); LC-MS (retention time: 1.49 min.), MS m/z 656(MH⁺).

Example 56 Preparation of Compound 56

Compound 56 was prepared by the same method as Compound 55 with thefollowing modifications:

Modifications: Cyclopentanecarbonyl chloride was used as a startingmaterial to give Compound 56 (18.0 mg, 24% yield): ¹H NMR (CD₃OD, 500MHz) δ 1.00-1.03 (m, 1H), 1.05 (s, 9H), 1.06-1.10 (m, 2H), 1.24-1.27 (m,2H), 1.25-1.61 (m, 9H), 1.80-1.83 (m, 1H), 1.88 (dd, J=8.24, 5.49 Hz,1H), 2.22-2.31 (m, 2H), 2.58-2.65 (m, 2H), 2.93-2.98 (m, 1H), 3.92 (s,3H), 4.10 (dd, J=11.90, 3.66 Hz, 1H), 4.35 (d, J=11.91 Hz, 1H), 4.52(dd, J=10.38, 7.02 Hz, 1H), 4.65 (d, J=9.46 Hz, 1H), 4.80 (d, J=5.49 Hz,1H), 4.88 (d, J=5.19 Hz, 1H), 5.13 (dd, J=10.37, 1.83 Hz, 1H), 5.30 (dd,J=16.80, 1.22 Hz, 1H), 5.73-5.78 (m, 1H), 5.84 (t, J=4.27 Hz, 1H), 7.11(dd, J=9.16, 2.44 Hz, 1H), 7.19 (d, J=2.44 Hz, 1H), 7.25 (d, J=5.80 Hz,1H), 7.88 (d, J=6.10 Hz, 1H), 8.05 (d, J=9.16 Hz, 1H)); LC-MS (retentiontime: 1.71 min.), MS m/z 710 (MH⁺).

Example 57 Preparation of Compound 57

Compound 57 was prepared by the same method as Compound 55 with thefollowing modifications:

Modifications: 2-Ethylbutyryl chloride was used as a starting materialto give Compound 57 (20.7 mg, 27% yield): ¹H NMR (CD₃OD, 500 MHz) δ 0.66(t, J=7.32 Hz, 3H), 0.85 (t, J=7.32 Hz, 3H), 1.02-1.05 (m, 1H), 1.07 (s,9H), 1.10-1.12 (m, 1H), 1.24-1.33 (m, 4H), 1.36-1.39 (m, 1H), 1.43 (dd,J=9.46, 5.19 Hz, 1H), 1.48-1.51 (m, 1H), 1.88 (dd, J=8.24, 5.19 Hz, 1H),2.12-2.14 (m, 1H), 2.22 (q, J=8.85 Hz, 1H), 2.26-2.30 (m, 1H), 2.59 (dd,J=13.73, 6.71 Hz, 1H), 2.94-2.97 (m, 1H), 3.92 (s, 3H), 4.11 (dd,J=11.90, 3.66 Hz, 1H), 4.40 (d, J=11.90 Hz, 1H), 4.50 (dd, J=10.68, 7.02Hz, 1H), 4.75 (d, J=9.46 Hz, 1H), 4.81 (d, J=9.16 Hz, 1H), 4.89 (d,J=9.16 Hz, 1H), 5.12 (dd, J=10.38, 1.53 Hz, 1H), 5.29 (dd, J=17.09, 1.22Hz, 1H), 5.72-5.79 (m, 1H), 5.85 (t, J=3.66 Hz, 1H), 7.08 (dd, J=9.16,2.44 Hz, 1H), 7.19 (d, J=2.44 Hz, 1H), 7.25 (d, J=5.80 Hz, 1H), 7.88 (d,J=6.10 Hz, 1H), 7.98 (d, J=9.16 Hz, 1H), 8.02 (d, J=9.16 Hz, 1H); LC-MS(retention time: 1.73 min.), MS m/z 712 (MH⁺).

Example 58 Preparation of Compound 58

To a solution mixture of product from step 1 of example 55 (70.0 mg,0.108 mmol), DIEA (41.8 mg, 0.323 mmol) and cyclopropaneacetic acid(16.2 mg, 0.162 mmol) in DCM (2 mL) was added HATU (61.6 mg, 0.162mmol). After stirring the reaction mixture at rt overnight, it waswashed with 5% aqueous NaHCO₃ (1 mL). The aqueous layer was extractedwith 2×2 mL DCM. The combined organic layer was washed with 5% aqueouscitric acid (2 mL), brine, dried over MgSO₄ and concentrated. Productwas purified by reversed phase prep-HPLC to give Compound 58 (21.9 mg,29% yield): ¹H NMR (CD₃OD, 500 MHz) δ 0.11-0.14 (m, 2H), 0.43-0.47 (m,2H), 0.87-0.09 (m, 1H), 1.01-1.04 (m, 1H), 1.07 (s, 9H), 1.09-1.12 (m,1H), 1.23-1.27 (m, 2H), 1.45 (dd, J=9.46, 5.49 Hz, 1H), 1.88 (dd,J=8.24, 5.49 Hz, 1H), 2.03 (d, J=7.32 Hz, 2H), 2.23 (q, J=8.75 Hz, 1H),2.27-2.31 (m, 1H), 2.59 (dd, J=13.73, 7.02 Hz, 1H), 2.92-2.96 (m, 1H),3.93 (m, 3H), 4.13 (dd, J=11.90, 3.97 Hz, 1H), 4.34 (d, J=11.90 Hz, 1H),4.53 (dd, J=10.38, 7.02 Hz, 1H), 4.66 (d, J=9.46 Hz, 1H), 4.81 (d,J=6.10 Hz, 1H), 4.89 (d, J=6.10 Hz, 1H), 5.12 (dd, J=10.37, 1.52 Hz,1H), 5.30 (dd, J=17.09, 1.22 Hz, 1H), 5.75-5.81 (m, 1H), 5.86 (s, 1H),7.12 (dd, J=9.16, 2.44 Hz, 1H), 7.19 (d, J=2.44 Hz, 1H), 7.81 (d, J=9.46Hz, 1H), 7.89 (d, J=5.80 Hz, 1H), 8.06 (d, J=9.16 Hz, 1H); LC-MS(retention time: 1.63 min.), MS m/z 696 (MH⁺).

Example 59 Preparation of Compound 59

Compound 59 was prepared by the same method as Compound 58 with thefollowing modifications:

Modifications: Methoxyacetic acid was used as a starting material togive Compound 59 (23.5 mg, 32% yield): ¹HNMR (CD₃OD, 500 MHz) δ10.99-1.04 (m, 2H), 1.06 (s, 9H), 1.09-1.12 (m, 1H), 1.22-1.27 (m, 2H),1.45 (dd, J=9.46, 5.49 Hz, 1H), 1.88 (dd, J=8.24, 5.49 Hz, 1H), 2.22 (q,J=8.85 Hz, 1H), 2.29-2.32 (m, 1H), 2.60 (dd, J=13.89, 6.87 Hz, 1H),2.92-2.97 (m, 1H), 3.35 (s, 3H), 3.70 (d, J=15.26 Hz, 1H), 3.84 (d,J=15.26 Hz, 1H), 3.93 (s, 3H), 4.13 (dd, J=11.90, 3.97 Hz, 1H), 4.32 (d,J=11.60 Hz, 1H), 4.54 (dd, J=10.38, 7.02 Hz, 1H), 4.65 (s, 1H), 4.81 (d,J=7.32 Hz, 1H), 4.89 (d, J=7.32 Hz, 1H), 5.12 (d, J=10.38 Hz, 1H), 5.30(d, J=16.79 Hz, 1H), 5.74-5.81 (m, 1H), 5.86 (t, J=3.36 Hz, 1H), 7.14(dd, J=9.00, 2.59 Hz, 1H), 7.19 (d, J=2.44 Hz, 1H), 7.26 (d, J=6.10 Hz,1H), 7.89 (d, J=6.10 Hz, 1H), 8.04 (d, J=9.16 Hz, 1H); LC-MS (retentiontime: 1.54 min.), MS m/z 686 (MH⁺).

Example 60 Preparation of Compound 60

Compound 60 was prepared by the same method as Compound 58 with thefollowing modifications:

Modifications: (+)-Methoxyacetic acid was used as a starting material togive Compound 60 (23.8 mg, 27% yield): ¹HNMR (CD₃OD, 500 MHz) δ 0.78 (d,J=7.02 Hz, 3H), 0.83-0.88 (m, 2H), 0.92 (t, J=7.17 Hz, 6H), 0.94-0.98(m, 1H), 1.00-1.03 (m, 2H), 1.06 (s, 9H), 1.07-1.11 (m, 1H), 1.22-1.26(m, 2H), 1.31-1.36 (m, 2H), 1.45 (dd, J=9.46, 5.49 Hz, 1H), 1.63-1.68(m, 2H), 1.89 (dd, J=8.09, 5.34 Hz, 1H), 2.01-2.04 (m, 1H), 2.17-2.21(m, 1H), 2.24 (q, J=9.00 Hz, 2H), 2.28-2.33 (m, 1H), 2.60 (dd, J=13.73,7.02 Hz, 1H), 2.93-2.98 (m, 1H), 3.15-3.20 (m, 1H), 3.77 (d, J=15.26 Hz,1H), 3.87 (d, J=15.26 Hz, 1H), 3.93 (s, 3H), 4.12 (dd, J=11.90, 3.66 Hz,1H), 4.32 (d, J=11.90 Hz, 1H), 4.56 (dd, J=10.38, 7.02 Hz, 1H), 4.65 (d,J=9.77 Hz, 1H), 4.81 (d, J=5.80 Hz, 1H), 4.89 (d, J=5.80 Hz, 1H), 5.13(dd, J=10.22, 1.68 Hz, 1H), 5.30 (dd, J=17.09, 1.53 Hz, 1H), 5.75-5.79(m, 1H), 5.85 (t, J=3.66 Hz, 1H), 7.14 (dd, J=9.16, 2.44 Hz, 1H), 7.19(d, J=2.44 Hz, 1H), 7.26 (d, J=5.80 Hz, 1H), 7.52 (d, J=9.77 Hz, 1H),7.89 (d, J=5.80 Hz, 1H), 8.03 (d, J=9.16 Hz, 1H); LC-MS (retention time:2.043 min.), MS m/z 810 (MH⁺).

Example 61 Preparation of Compound 61

Compound 61 was prepared by the same method as Compound 58 with thefollowing modifications:

Modifications: (−)-Methoxyacetic acid was used as a starting material togive Compound 61 (26.4 mg, 30% yield): ¹HNMR (CD₃OD, 500 MHz) δ 0.78 (d,J=7.02 Hz, 3H), 0.82-084 (m, 1H), 0.88 (dd, J=8.39, 3.81 Hz, 1H), 0.91(d, J=7.01 Hz, 3H), 0.92 (d, J=6.41 Hz, 3H), 0.94-0.99 (m, 2H),1.00-1.03 (m, 2H), 1.06 (s, 9H), 1.08-1.10 (m, 1H), 1.23-1.26 (m, 2H),1.30-1.37 (m, 2H), 1.44 (dd, J=9.61, 5.34 Hz, 1H), 1.62-1.68 (m, 2H),1.89 (dd, J=8.24, 5.49 Hz, 1H), 1.98-2.02 (m, 1H), 2.13-2.16 (m, 1H),2.24 (q, J=8.85 Hz, 1H), 2.28-2.32 (m, 1H), 2.60 (dd, J=13.73, 7.02 Hz,1H), 2.94-2.98 (m, 1H), 3.08-3.13 (m, 1H), 3.63 (d, J=15.56 Hz, 1H),3.93 (S, 3H), 4.11 (dd, J=12.05, 3.81 Hz, 1H), 4.32 (d, J=11.90 Hz, 1H),4.56 (dd, J=10.38, 7.02 Hz, 1H), 4.62 (d, J=9.46 Hz, 1H), 4.81 (d,J=6.41 Hz, 1H), 4.89 (d, J=6.72 Hz, 1H), 5.13 (dd, J=10.38, 1.83 Hz,1H), 5.30 (dd, J=17.09, 1.23 Hz, 1H), 5.76-5.80 (m, 1H), 5.85 (t, J=3.51Hz, 1H), 7.14 (dd, J=9.00, 2.59 Hz, 1H), 7.20 (d, J=2.44 Hz, 1H), 7.26(d, J=5.80 Hz, 1H), 7.52 (d, J=9.77 Hz, 1H), 7.89 (d, J=6.10 Hz, 1H),8.04 (d, J=9.16 Hz, 1H); LC-MS (retention time: 2.05 min.), MS m/z 810(MH⁺).

Example 62 Preparation of Compound 62

Compound 62 was prepared by the same method as Compound 58 with thefollowing modifications:

Modifications: Bicyclo[1.1.1]pentane-2-carboxylic acid was used as astarting material to give Compound 62 (35.1, 45% yield): ¹HNMR (CD₃OD,500 MHz) δ 1.03 (d, J=5.49 Hz, 1H), 1.06 (s, 9H), 1.08-1.11 (m, 3H),1.23-1.29 (m, 3H), 1.37 (dd, J=7.02, 3.36 Hz, 13H), 1.46 (dd, J=9.46,5.19 Hz, 1H), 1.65 (dd, J=9.77, 2.14 Hz, 1H), 1.69 (d, J=2.14 Hz, 1H),1.72 (dd, J=7.32, 3.05 Hz, 1H), 1.88 (dd, J=8.09, 5.34 Hz, 1H), 2.12(dd, J=9.77, 3.05 Hz, 1H), 2.23 (d, J=8.85 Hz, 1H), 2.27-2.31 (m, 1H),2.60 (t, J=6.87 Hz, 1H), 2.63 (d, J=1.83 Hz, 2H), 2.68 (d, J=7.63 Hz,1H), 2.93-2.96 (m, 1H), 3.23 (q, J=7.43 Hz, 2H), 3.70-3.75 (m, 2H), 3.93(s, 3H), 4.11 (dd, J=11.90, 3.66 Hz, 1H), 4.35 (d, J=11.90 Hz, 1H), 4.53(dd, J=10.53, 6.87 Hz, 1H), 4.71 (d, J=9.46 Hz, 1H), 4.79 (d, J=5.80 Hz,2H), 4.87 (d, J=5.49 Hz, 2H), 5.13 (dd, J=10.38, 1.83 Hz, 1H), 5.30 (dd,J=17.24, 1.37 Hz, 1H), 5.74-5.79 (m, 1H), 5.86 (t, J=3.20 Hz, 1H), 7.12(dd, J=9.16, 2.44 Hz, 1H), 7.19 (d, J=2.44 Hz, 1H), 7.25 (d, J=5.80 Hz,1H), 7.72 (d, J=9.46 Hz, 1H), 7.88 (d, J=5.80 Hz, 1H), 8.05 (d, J=9.16Hz, 1H); LC-MS (retention time: 1.16 min.), MS m/z 708 (MH⁺).

Example 63 Preparation of Compound 63

Compound 63 was prepared by the same method as Compound 58 with thefollowing modifications:

Modifications: pyrazine-2-carboxylic acid was used as a startingmaterial to give Compound 63 (42.3, 54% yield): ¹HNMR (CD₃OD, 500 MHz) δ1.00-1.04 (m, 3H), 1.05-1.09 (m, 1H), 1.10 (s, 9H), 1.15 (s, 1H),1.18-1.22 (m, 2H), 1.43 (dd, J=9.46, 5.49 Hz, 1H), 1.88 (dd, J=7.93,5.49 Hz, 1H), 2.17 (q, J=8.65 Hz, 1H), 2.37-2.42 (m, 1H), 2.64 (dd,J=13.73, 7.32 Hz, 1H), 2.91-2.95 (m, 1H), 3.88 (s, 3H), 3.93 (d, J=3.35Hz, 1H), 4.13 (dd, J=11.90, 3.36 Hz, 1H), 4.46 (d, J=11.90 Hz, 1H), 4.61(dd, J=10.07, 7.32 Hz, 1H), 4.76 (s, 1H), 4.80 (d, J=7.63 Hz, 1H), 4.88(d, J=7.93 Hz, 1H), 5.09 (d, J=10.38 Hz, 1H), 5.27 (d, J=17.09 Hz, 1H),5.77-5.83 (m, 1H), 5.85 (s, 1H), 6.85 (dd, J=9.16, 2.44 Hz, 1H), 7.08(d, J=2.14 Hz, 1H), 7.23 (d, J=5.80 Hz, 1H), 7.87 (d, J=8.85 Hz, 1H),7.90 (d, J=6.10 Hz, 1H), 8.57 (d, J=1.53 Hz, 1H), 8.73 (d, J=2.44 Hz,1H), 8.81 (s, 1H).

Example 64 Preparation of Compound 64

To a solution mixture of product from step 1 of example 55 (70.0 mg,0.108 mmol) and DIEA (41.8 mg, 0.323 mmol) in DCM (2 mL) was addedbenzyl chloroformate (55.1 mg, 0.323 mmol). After stirring at rt for 14h, solvent was removed and product was purified by reversed phaseprep-HPLC to give Compound 64 (26.9 mg, 31% yield): ¹HNMR (CD₃OD, 500MHz) δ 1.00 (d, J=2.14 Hz, 1H), 1.02 (d, J=5.80 Hz, 1H), 1.04 (s, 9H),1.08-1.14 (m, 1H), 1.16 (d, J=6.71 Hz, 1H), 1.18-1.22 (m, 2H), 1.43 (dd,J=9.46, 5.19 Hz, 1H), 1.87 (dd, J=8.09, 5.34 Hz, 1H), 2.17-2.22 (m, 1H),2.30-2.35 (m, 1H), 2.62 (dd, J=13.73, 7.02 Hz, 1H), 2.90-2.95 (m, 1H),3.88 (s, 3H), 4.08 (dd, J=11.90, 3.66 Hz, 1H), 4.31 (s, 1H), 4.43 (d,J=11.60 Hz, 1H), 4.55 (dd, J=10.07, 7.32 Hz, 1H), 4.74 (d, J=12.21 Hz,1H), 4.81 (d, J=6.10 Hz, 1H), 4.89 (d, J=5.79 Hz, 1H), 5.10 (d, J=9.16Hz, 1H), 5.16 (s, 1H), 5.28 (d, J=17.09 Hz, 1H), 5.75-5.81 (m, 1H), 5.83(s, 1H), 7.07 (dd, J=9.16, 2.44 Hz, 1H), 7.17 (d, J=2.44 Hz, 1H), 7.20(d, J=7.32 Hz, 2H), 7.25 (t, J=5.65 Hz, 3H), 7.30-7.33 (m, 1H),7.34-7.37 (m, 2H), 7.89 (d, J=5.80 Hz, 1H), 8.07 (d, J=9.16 Hz, 1H);LC-MS (retention time: 1.79 min.), MS m/z 748 (MH⁺).

Example 65 Preparation of Compound 65

Compound 65 was prepared by the same method as Compound 64 with thefollowing modifications:

Modifications: (+)-Methyl chloroformate was used as a starting materialto give Compound 65 (28.8 mg, 36% yield): ¹H NMR (CD₃OD, 500 MHz) δ 0.72(d, J=6.71 Hz, 3H), 0.80 (t, J=5.80 Hz, 6H), 0.87 (d, J=7.02 Hz, 4H),0.90-0.95 (m, 6H), 0.98-1.02 (m, 5H), 1.05 (s, 9H), 1.07-1.12 (m, 2H),1.18-1.23 (m, 2H), 1.32-1.38 (m, 3H), 1.41 (dd, J=9.46, 5.19 Hz, 1H),1.46-1.48 (m, 1H), 1.63-1.71 (m, 5H), 1.85 (dd, J=7.93, 5.49 Hz, 1H),1.89-1.93 (m, 1H), 2.00-2.03 (m, 1H), 2.15 (q, J=8.70 Hz, 1H), 2.34-2.38(m, 1H), 2.61 (dd, J=13.73, 7.33 Hz, 1H), 2.89-2.93 (m, 1H), 3.73 (s,2H), 3.92 (s, 3H), 4.10 (dd, J=11.60, 3.36 Hz, 1H), 4.33 (s, 1H), 4.41(d, J=11.29 Hz, 1H), 4.46-4.52 (m, 1H), 4.54 (dd, J=9.76, 7.90 Hz, 1H),4.81 (d, J=5.80 Hz, 1H), 4.89 (m, 1H), 5.08 (d, J=11.60 Hz, 1H), 5.26(d, J=17.09 Hz, 1H), 5.77-5.81 (m, 1H), 5.83 (d, J=3.97 Hz, 1H), 7.11(dd, J=11.29, 1.83 Hz, 1H), 7.18 (d, J=1.83 Hz, 1H), 7.24 (d, J=5.80 Hz,1H), 7.88 (d, J=6.10 Hz, 1H), 8.08 (d, J=8.85 Hz, 1H); LC-MS (retentiontime: 2.06 min.), MS m/z 796 (MH⁺).

Example 66 Preparation of Compound 66

Compound 66 was prepared by the same method as Compound 64 with thefollowing modifications:

Modifications: (−)-Methyl chloroformate was used as a starting materialto give Compound 66 (26.9 mg, 31% yield): ¹H NMR (CD₃OD, 500 MHz) δ 0.35(d, J=6.41 Hz, 1H), 0.51 (d, J=6.71 Hz, 2H), 0.68 (d, J=6.71 Hz, 1H),0.73 (d, J=7.02 Hz, 2H), 0.77-0.82 (m, 4H), 0.88-0.98 (m, 10H), 1.0-1.03(m, 2H), 1.05 (s, 9H), 1.09-1.18 (m, 3H), 1.25-1.29 (m, 1H), 1.3-1.41(m, 3H), 1.60-1.71 (m, 3H), 1.82-1.89 (m, 3H), 2.00-2.04 (m, J=2.14 Hz,1H), 2.10 (q, J=8.24 Hz, 1H), 2.39-2.43 (m, 1H), 2.61 (dd, J=14.04, 7.32Hz, 1H), 2.87-2.91 (m, 1H), 3.73 (s, 1H), 3.92 (s, 3H), 4.13 (dd,J=11.75, 3.51 Hz, 1H), 4.22-4.27 (m, 2H), 4.30 (s, 1H), 4.39 (d, J=11.90Hz, 1H), 4.48-4.55 (m, 1H), 4.79 (d, J=5.19 Hz, 1H), 4.87 (d, J=4.27 Hz,1H), 5.05 (d, J=10.07 Hz, 1H), 5.22 (d, J=16.79 Hz, 1H), 5.78-5.85 (m,2H), 7.09 (dd, J=9.16, 1.83 Hz, 1H), 7.17 (d, J=1.83 Hz, 1H), 7.23 (d,J=5.80 Hz, 1H), 7.88 (d, J=5.80 Hz, 1H) 8.09 (d, J=9.16 Hz, 1H); LC-MS(retention time: 2.05 min.), MS m/z 796 (MH⁺).

Example 67 Preparation of Compound 67

Compound 67 was prepared by the same method as Compound 64 with thefollowing modifications:

Modifications: Di-tert-amyl dicarbonate was used as a starting materialto give Compound 67 (35.3 mg, 41% yield): ¹H NMR (CD₃OD, 500 MHz) δ 0.78(t, J=7.17 Hz, 3H), 0.89-0.95 (m, 6H), 1.04 (s, 9H), 1.005-1.09 (m, 3H),1.15 (s, 1H), 1.22 (d, J=12.51 Hz, 6H), 1.40 (s, 2H), 1.42-1.46 (m, 1H),1.48 (s, 3H), 1.56-1.66 (m, 2H), 1.78 (q, J=7.63 Hz, 1H), 1.84 (q,J=7.53 Hz, 1H), 1.88 (d, J=5.80 Hz, 1H), 2.22 (d, J=8.55 Hz, 1H),2.27-2.31 (m, 1H), 2.61 (dd, J=13.73, 7.02 Hz, 1H), 2.91-2.96 (m, 1H),3.92 (s, 3H), 4.08 (d, J=12.21 Hz, 1H), 4.26 (d, J=9.16 Hz, 1H), 4.42(d, J=11.29 Hz, 1H), 4.52 (t, J=7.93 Hz, 1H), 5.12 (d, J=10.07 Hz, 1H),5.29 (d, J=17.09 Hz, 1H), 5.73-5.80 (m, 1H), 5.84 (s, 1H), 6.57 (d,J=8.85 Hz, 1H), 7.09 (d, J=8.54 Hz, 1H), 7.18 (s, 1H), 7.25 (d, J=5.80Hz, 1H), 7.89 (d, J=5.80 Hz, 1H), 8.08 (d, J=9.16 Hz, 1H); LC-MS(retention time: 1.82 min.), MS m/z 728 (MH⁺).

Example 68 Preparation of Compound 68

Compound 68 was prepared by the same method as Compound 64 with thefollowing modifications:

Modifications: 2,2,2-Trichloro-1,1-dimethyl chloroformate was used as astarting material to give Compound 68 (30.5 mg, 37% yield): ¹H NMR(CD₃OD, 500 MHz) δ 0.99 (s, 9H), 1.04 (s, 6H), 1.08-1.09 (m, 3H),1.23-1.26 (m, 3H), 1.44 (s, 2H), 1.46 (d, J=5.80 Hz, 1H), 1.71 (s, 2H),2.23-2.33 (m, 2H), 2.60-2.64 (m, 1H), 2.93-2.96 (m, 1H), 3.70 (m, 1H),3.71 (s, 3H), 3.93 (s, 3H), 4.04-4.06 (m, 2H), 4.27 (d, J=9.16 Hz, 1H),4.41 (d, J=11.60 Hz, 1H), 4.57 (d, J=10.98, 6.11 Hz, 1H), 5.14 (d,J=12.21 Hz, 1H), 5.32 (d, J=17.70 Hz, 1H), 5.75-5.80 (m, 1H), 5.84 (s,1H), 7.10 (dd, J=9.16, 2.44 Hz, 1H), 7.19 (d, J=2.44 Hz, 1H), 7.26 (d,J=6.10 Hz, 1H), 7.90 (d, J=5.80 Hz, 1H), 8.07 (d, J=9.16 Hz, 1H)); LC-MS(retention time: 1.95 min.), MS m/z 816 (MH⁺).

Example 69 Preparation of Compound 69

To a solution mixture of product from step 1 of example 55 (102 mg,0.149 mmol) and DIEA (48.2, 0.373 mmol) in THF (2 mL) was addedN,N′-dissucinimidyl carbonate (57.1 mg, 0.223 mmol). The resultingsuspension was irradiated in a microwave to 80° C. for 15 min. Then wasadded a slurry solution of sodium 1-methyl cyclopentoxide which wasprepared by treating a 0° C. solution of 1-methyl cyclopentanol (149.2mg, 1.49 mmol) in THF (1 mL) with NaH (60% in oil, 59.6 mg, 1.49 mmol)for 15 min at rt. After stirring at rt 15 min, the reaction was quenchedwith saturated aqueous ammonium chloride (3 mL) and extracted with EtOAc(10 mL). The organic layer was then passed through a celite hydromatrixcolumn, concentrated and purified by reversed phase prep-HPLC to giveCompound 69 (49.0 mg, 44%): ¹H NMR (CD₃OD, 500 MHz) δ 0.95-0.98 (m, 3H),0.99-1.01 (m, J=12.51 Hz, 1H), 1.03 (s, 9H), 1.14-1.18 (m, 2H), 1.30 (s,3H), 1.40-1.47 (m, 3H), 1.50-1.56 (m, 3H), 1.60-1.64 (m, 1H), 1.76-1.81(m, 1H), 1.83-1.85 (m, 1H), 2.10-2.19 (m, 1H), 2.36-2.43 (m, 1H), 2.63(dd, J=14.50, 7.17 Hz, 1H), 2.86-2.90 (m, 1H), 3.92 (s, 3H), 4.09 (d,J=12.51 Hz, 1H), 4.25 (d, J=1.53 Hz, 1H), 4.43 (d, J=10.99 Hz, 1H),4.51-4.55 (m, 1H), 5.06 (d, J=11.60 Hz, 1H), 5.23 (d, J=16.78 Hz, 1H),5.80-5.85 (m, J=12.67, 12.67 Hz, 2H), 7.09 (d, J=8.55 Hz, 1H), 7.17 (s,1H), 7.24 (d, J=5.49 Hz, 1H), 8.07 (d, J=9.16 Hz, 1H)); LC-MS (retentiontime: 1.87 min.), MS m/z 740 (MH⁺).

Example 70 Preparation of Compound 70

Compound 70 was prepared by the same method as Compound 69 with thefollowing modifications:

Modifications: Cyclopentanol was used as a starting material to giveCompound 70 (85.1 mg, 40% yield): ¹H NMR (CD₃OD, 500 MHz) δ 0.98 (s,1H), 1.00 (d, J=4.88 Hz, 1H), 1.03 (s, 9H), 1.06-110 (m, 2H), 1.24-1.29(m, 3H), 1.36-1.40 (m, 2H), 1.44 (dd, J=9.31, 5.04 Hz, 2H), 1.57-1.62(m, 5H), 1.69-1.73 (m, 2H), 1.88 (dd, J=8.09, 5.65 Hz, 1H), 2.22-29 (m,2H), 2.59-2.62 (m, 1H), 2.92-2.96 (m, 1H), 3.93 (s, 3H), 4.07 (dd,J=10.99, 2.44 Hz, 1H), 4.29 (s, 1H), 4.42 (dd, J=12.51, 1.53 Hz, 1H),4.55 (dd, J=9.77, 7.93 Hz, 1H), 4.68-4.71 (m, 1H), 4.81 (d, J=8.55 Hz,1H), 4.89 (d, J=9.46 Hz, 1H), 5.13 (d, J=10.68 Hz, 1H), 5.30 (d, J=16.48Hz, 1H), 5.73-5.78 (m, 1H), 5.84 (s, 1H), 7.12 (dd, J=9.15, 1.83 Hz,1H), 7.20 (d, J=2.14 Hz, 1H), 7.27 (d, J=5.80 Hz, 1H), 7.89 (d, J=5.80Hz, 1H), 8.09 (d, J=8.85 Hz, 1H); LC-MS (retention time: 1.81 min.), MSm/z 726 (MH⁺).

Example 71 Preparation of Compound 71

Compound 71 was prepared by the same method as Compound 69 with thefollowing modifications:

Modifications: Cyclobutanol was used as a starting material to giveCompound 71 (16.2 mg, 39% yield): ¹H NMR (CD₃OD, 500 MHz) δ 0.97 (s,3H), 1.03 (s, 9H), 1.06-1.08 (m, 2H), 1.17-1.22 (m, 3H), 1.37-1.44 (m,1H), 1.82-1.85 (m, 1H), 2.02-2.08 (m, 1H), 2.15-2.21 (m, 1H), 2.30-2.36(m, 1H), 2.49-2.54 (m, 0.4H), 2.59-2.67 (m, 0.6H), 2.90-2.94 (m, 1H),3.93 (s, 3H), 4.09 (dd, J=8.09, 4.73 Hz, 1H), 4.20 (d, J=10.68 Hz,0.4H), 4.38 (d, J=10.68 Hz, 0.6H), 4.46 (dd, J=10.22, 7.17 Hz, 0.4H),4.48-4.56 (m, 1H), 5.07-5.11 (m, 1H), 5.26 (d, J=17.24 Hz, 0.4H), 5.28(d, J=17.24 Hz, 0.6H), 5.71-5.79 (m, 1H), 5.84 (s, 1H), 7.07 (dd,J=8.39, 2.90 Hz, 1H), 7.14 (d, J=2.14 Hz, 1H), 7.20 (d, J=2.44 Hz, 1H),7.25 (d, J=5.80 Hz, 1H), 7.59 (d, J=5.80 Hz, 1H), 7.88 (m, 1H), 8.00 (d,J=9.16 Hz, 0.4H), 8.07 (d, J=8.85 Hz, 0.6H)); LC-MS (retention time:1.25 min.), MS m/z 712 (MH⁺).

Example 72 Preparation of Compound 72

Compound 72 was prepared by the same method as Compound 69 with thefollowing modifications:

Modifications: 2-Phenyl-2-propanol was used as a starting material togive Compound 72 (19.0 mg, 42% yield): ¹H NMR (CD₃OD, 500 MHz) δ 0.97(m, 1H), 1.03 (s, 9H), 1.06-1.09 (m, 3H), 1.16-1.22 (m, 4H), 1.41-1.44(m, 1H), 1.57 (s, 3H), 1.86 (t, J=7.80 Hz, 1H), 2.14-2.18 (m, 1H),2.30-2.35 (m, 1H), 2.57-2.61 (m, 1H), 2.90-2.94 (m, 1H), 3.92 (d, J=4.27Hz, 1H), 3.94 (s, 3H), 4.04 (dd, J=10.99, 3.66 Hz, 1H), 4.18 (s, 1H),4.24 (d, J=10.99 Hz, 1H), 4.52 (s, 1H), 5.09 (d, J=10.07 Hz, 1H), 5.26(d, J=14.95 Hz, 1H), 5.78-5.82 (m, 2H), 7.07-7.12 (m, 2H), 7.16-7.20 (m,3H), 7.23 (d, J=5.19 Hz, 1H), 7.29 (d, J=7.02 Hz, 2H), 7.84 (d, J=5.80Hz, 1H), 8.03 (d, J=9.46 Hz, 1H)); LC-MS (retention time: 1.84 min.), MSm/z 776 (MH⁺).

Example 73 Preparation of Compound 73

Compound 73 was prepared by the same method as Compound 69 with thefollowing modifications:

Modifications: 4-(Trifluoromethyl)phenyl dimethyl carbonyl was used as astarting material to give Compound 73 (22.1 mg, 45% yield): ¹H NMR(CD₃OD, 500 MHz) δ 0.91 (s, 1H), 0.97-1.00 (m, J=15.56 Hz, 4H), 1.04 (s,9H), 1.07-1.10 (m, 2H), 1.16-1.20 (m, 3H), 1.30-1.31 (m, 1H), 1.41 (dd,J=9.61, 5.34 Hz, 1H), 1.55 (d, J=7.32 Hz, 6H), 1.83-1.87 (m, 1H),2.11-2.14 (m, 1H), 2.34-2.39 (m, 1H), 2.57-2.62 (m, 1H), 2.89-2.92 (m,J=11.60, 4.27 Hz, 1H), 3.92 (s, 2H), 3.94 (s, 3H), 4.02-4.05 (m, 1H),4.17 (s, 1H), 4.26 (d, J=11.90 Hz, 1H), 4.53 (t, J=8.85 Hz, 1H), 5.07(d, J=10.07 Hz, 1H), 5.24 (d, J=18.01 Hz, 1H), 5.78-5.83 (m, 2H), 7.08(d, J=7.02 Hz, 1H), 7.19 (s, 1H), 7.22 (d, J=5.80 Hz, 1H), 7.45 (dd,J=13.74, 7.63 Hz, 3H), 7.60 (d, J=6.41 Hz, 1H), 7.67 (d, J=7.63 Hz, 1H),7.84 (d, J=5.80 Hz, 1H), 8.02 (d, J=8.54 Hz, 1H)); LC-MS (retentiontime: 1.92 min.), MS m/z 844 (MH⁺).

Example 74 Preparation of Compound 74

To a solution mixture of the product from step 1 of example 55 (70.0 mg,0.108 mmol) and DIEA (41.8 mg, 0.323 mmol) in DCM (2 mL) was addedt-butylisocyanate (32.0, 0.323 mmol). After stirring at rt overnight,the reaction was concentrated and purified by reversed-phase prep-HPLCto give Compound 74 (42.3 mg, 55% yield): ¹H NMR (CD₃OD, 500 MHz) δ0.96-1.00 (m, 1H) 1.04 (s, 9H) 1.08-1.10 (m, 3H) 1.19 (s, 9H) 1.22-1.31(m, 2H) 1.30 (m, 1H) 1.41 (dd, J=9.46, 5.49 Hz, 1H) 1.87 (dd, J=8.24,5.49 Hz, 1H) 2.20-2.29 (m, 2H) 2.61 (dd, J=14.04, 6.72 Hz, 1H) 2.92-2.97(m, 1H) 3.92 (s, 3H) 4.08 (dd, J=11.60, 3.97 Hz, 1H) 4.36 (s, 1H)4.47-4.52 (m, 2H) 4.81 (d, J=3.36 Hz, 1H) 4.88 (d, J=8.85 Hz, 1H) 5.11(dd, J=10.22, 1.68 Hz, 1H) 5.28 (dd, J=17.09, 1.53 Hz, 1H) 5.72-5.76 (m,1H) 5.85 (s, 1H) 7.08 (dd, J=9.16, 2.44 Hz, 1H) 7.18 (d, J=2.14 Hz, 1H)7.24 (d, J=5.80 Hz, 1H) 7.88 (d, J=6.10 Hz, 1H) 8.12 (d, J=9.16 Hz, 1H);LC-MS (retention time: 1.70 min.), MS m/z 713 (MH⁺).

Example 75 Preparation of Compound 75

Compound 75 was prepared by the same method as Compound 74 with thefollowing modifications:

Modifications: Cyclopentyl isocyanate was used as a starting material togive Compound 75 (38.5 mg, 49%): ¹H NMR (CD₃OD, 500 MHz) δ 0.92 (d,J=7.63 Hz, 1H) 0.96 (s, 9H) 0.98-1.02 (m, 1H) 1.05 (s, 9H) 1.07-1.10 (m,2H) 1.21-1.25 (m, 3H) 1.28-1.34 (m, 1H) 1.36-1.55 (m, 8H) 1.58-1.65 (m,13H) 1.81 (m, 1H) 1.88 (m, 6H) 2.23 (dd, J=18.01, 8.85 Hz, 1H) 2.29 (m,1H) 2.59 (dd, J=13.73, 7.02 Hz, 1H) 2.94 (m, 1H) 3.27 (d, J=1.83 Hz, 1H)3.35 (d, J=1.53 Hz, 1H) 3.75 (m, 1H) 3.92 (s, 3H) 3.95 (d, J=6.41 Hz,1H) 3.97 (s, 1H) 4.09 (m, 2H) 4.40 (s, 1H) 4.45 (d, J=11.90 Hz, 1H) 4.52(dd, J=10.07, 7.02 Hz, 1H) 4.81 (d, J=7.02 Hz, 1H) 4.89 (d, J=7.02 Hz,1H) 5.11 (m, 1H) 5.29 (d, J=17.40 Hz, 1H) 5.75 (m, 1H) 5.85 (s, 1H) 7.11(dd, J=9.16, 2.44 Hz, 1H) 7.18 (d, J=2.44 Hz, 1H) 7.25 (d, J=5.80 Hz,1H) 7.88 (d, J=6.10 Hz, 1H) 7.95 (m, 1H) 8.12 (d, J=9.16 Hz, 1H)); LC-MS(retention time: 1.67 min.), MS m/z 725 (MH⁺).

Example 76 Preparation of Compound 76

To a solution mixture of the product from step 1 of example 55 (70 mg,0.102 mmol) and DIEA (33.0 mg, 0.255 mmol) in THF (2 mL) was addedN,N′-dissucinimidyl carbonate (39.2 mg, 0.153 mmol). The resultingsuspension was irradiated in a microwave to 80° C. for 15 min. Then itwas treated with tert-amylamine (88.9 mg, 1.02 mmol). After stirring atrt 15 min, the reaction was concentrated and purified by reversed phaseprep-HPLC to give Compound 76 (51 mg, 69%): ¹H NMR (CD₃OD, 500 MHz) δ0.76 (t, J=7.48 Hz, 4H), 0.97 (s, 1H), 1.04 (s, 9H), 1.13 (s, 6H),1.22-1.25 (m, 2H), 1.41 (dd, J=9.61, 5.34 Hz, 1H), 1.53 (dd, J=13.89,7.48 Hz, 1H), 1.58-1.62 (m, 1H), 1.87 (dd, J=7.93, 5.49 Hz, 1H), 2.20(q, J=8.65 Hz, 1H), 2.27-2.31 (m, 1H), 2.60 (dd, J=13.73, 7.32 Hz, 1H),2.92-2.96 (m, 1H), 3.92 (s, 3H), 4.08 (dd, J=11.75, 3.81 Hz, 1H), 4.36(s, 1H), 4.46 (d, J=11.90 Hz, 1H), 4.50 (dd, J=10.22, 7.17 Hz, 1H), 5.10(dd, J=10.22, 1.37 Hz, 1H), 5.27 (dd, J=16.94, 1.07 Hz, 1H), 5.73-5.77(m, 1H), 5.84 (t, J=3.51 Hz, 1H), 7.09 (dd, J=9.16, 2.44 Hz, 1H), 7.18(d, J=2.44 Hz, 1H), 7.24 (d, J=5.80 Hz, 1H), 7.88 (d, J=6.10 Hz, 1H),8.11 (d, J=9.16 Hz, 1H); LC-MS (retention time: 1.753 min.), MS m/z 727(MH⁺).

Example 77 Preparation of Compound 77

Compound 77 was prepared by the same method as Compound 76 with thefollowing modifications:

Modifications: tert-Butyl methylamine was used as a starting material togive Compound 77 (160.7 mg, 74% yield): ¹HNMR (CD₃OD, 500 MHz) δ0.96-1.10 (m, 1H), 1.06 (s, 9H), 1.08-1.12 (m, J=5.80 Hz, 3H), 1.26 (s,9H), 1.46 (dd, J=9.46, 5.19 Hz, 1H), 1.87 (dd, J=7.93, 5.49 Hz, 1H),2.21 (q, J=8.75 Hz, 1H), 2.26-2.31 (m, 1H), 2.57-2.62 (m, 1H), 2.86 (s,3H), 2.91-2.95 (m, 1H), 3.92 (s, 3H), 4.09 (dd, J=11.90, 3.66 Hz, 1H),4.43 (s, 1H), 4.46 (d, J=11.90 Hz, 1H), 4.52 (dd, J=10.68, 7.02 Hz, 1H),5.11 (dd, J=10.22, 1.37 Hz, 1H), 5.29 (d, J=17.09 Hz, 1H), 5.75-5.82 (m,1H), 5.86 (s, 1H), 7.09 (dd, J=9.16, 2.14 Hz, 1H), 7.18 (d, J=2.44 Hz,1H), 7.24 (d, J=5.80 Hz, 1H), 7.88 (d, J=5.80 Hz, 1H), 8.09 (d, J=8.85Hz, 1H)); LC-MS (retention time: 1.76 min.), MS m/z 727 (MH⁺).

Example 78 Preparation of Compound 78

Compound 78 was prepared by the same method as Compound 76 with thefollowing modifications:

Modifications: N,O-Dimethylhydroxylamine hydrochloride was used as astarting material to give Compound 78 (62.1 mg, 60% yield): ¹HNMR(CD₃OD, 500 MHz) δ 0.99 (t, J=6.10 Hz, 1H), 1.07 (s, 11H), 1.22-1.26 (m,J=3.97 Hz, 2H), 1.47 (dd, J=9.46, 5.49 Hz, 1H), 1.88 (dd, J=8.24, 5.49Hz, 1H), 2.22 (d, J=8.54 Hz, 1H), 2.3-30-2.33 (m, 1H), 2.60 (dd,J=13.43, 7.02 Hz, 1H), 2.92 (s, 3H), 2.93-2.96 (m, 1H), 3.66 (s, 3H),3.92 (s, 3H), 4.12 (dd, J=11.90, 3.66 Hz, 1H), 4.34 (d, J=12.21 Hz, 1H),4.44 (d, J=9.46 Hz, 1H), 4.54 (dd, J=10.53, 6.87 Hz, 1H), 5.12 (d,J=10.38 Hz, 1H), 5.30 (d, J=17.09 Hz, 1H), 5.75-5.83 (m, 1H), 5.86 (t,J=3.97 Hz, 1H), 6.70 (d, J=9.77 Hz, 1H), 7.13 (dd, J=9.16, 2.44 Hz, 1H),7.19 (d, J=2.44 Hz, 1H), 7.25 (d, J=6.10 Hz, 1H), 7.88 (d, J=5.80 Hz,1H), 8.07 (d, J=9.16 Hz, 1H)); LC-MS (retention time: 1.59 min.), MS m/z701 (MH⁺).

Example 79 Preparation of Compound 79

Compound 79 was prepared by the same method as Compound 76 with thefollowing modifications:

Modifications: Diethylamine was used as a starting material to giveCompound 79 (56.5 mg, 54% yield): ¹HNMR (CD₃OD, 500 MHz) δ 1.03 (q,J=15.6 Hz, 4H), 1.06 (d, J=1.53 Hz, 9H), 1.05-1.10 (m, 3H), 1.13-1.23(m, 4H), 1.46 (dd, J=9.46, 5.19 Hz, 1H), 1.86 (dd, J=7.93, 5.30 Hz, 1H),2.17 (q, J=8.85 Hz, 1H), 2.32-2.36 (m, 1H), 2.60 (dd, J=14.04, 7.32 Hz,1H), 2.89-2.93 (m, 1H), 3.16-3.24 (m, 4H), 3.92 (s, 3H), 4.14 (dd,J=11.90, 3.66 Hz, 1H), 4.37 (d, J=11.60 Hz, 1H), 4.51-4.55 (m, 2H), 5.09(d, J=10.07 Hz, 1H), 5.27 (d, J=17.09 Hz, 1H), 5.55 (d, J=9.46 Hz, 1H),5.79-5.84 (m, 1H), 5.86 (s, 1H), 7.11 (dd, J=8.85, 2.44 Hz, 1H), 7.18(d, J=2.44 Hz, 1H), 7.24 (d, J=5.80 Hz, 1H), 7.88 (d, J=5.80 Hz, 1H),8.08 (d, J=9.16 Hz, 1H)); LC-MS (retention time: 1.68 min.), MS m/z 713(MH⁺).

Example 80 Preparation of Compound 80

Compound 80 was prepared by the same method as Compound 76 with thefollowing modifications:

Modifications: Saturated aqueous ammonium chloride was used as astarting material to give Compound 76 (12.2 mg, 32% yield): ¹HNMR(CD₃OD, 500 MHz) δ 1.00-1.03 (m, 3H), 1.06 (s, 9H), 1.20-1.25 (m, 2H),1.42 (dd, J=9.31, 5.34 Hz, 1H), 2.22 (d, J=9.77 Hz, 1H), 2.29-2.35 (m,1H), 2.59 (dd, J=13.28, 6.87 Hz, 1H), 2.92-2.96 (m, 1H), 3.92 (s, 3H),4.14 (dd, J=11.75, 4.12 Hz, 1H), 4.38-4.43 (m, 1H), 4.51 (dd, J=9.92,6.87 Hz, 1H), 5.11 (d, J=11.90 Hz, 1H), 5.28 (d, J=17.70 Hz, 1H),5.72-5.79 (m, 1H), 5.84 (s, 1H), 7.15 (d, J=2.44 Hz, 1H), 7.17 (d,J=2.75 Hz, 1H), 7.23 (d, J=5.80 Hz, 1H), 7.87 (d, J=7.87 Hz, 1H), 8.10(d, J=8.85 Hz, 1H)); LC-MS (retention time: 1.43 min.), MS m/z 657(MH⁺).

Example 81 Preparation of Compound 81

Compound 81 was prepared by the same method as Compound 76 with thefollowing modifications:

Modifications: tert-Octylamine was used as a starting material to giveCompound 81 (16.1 mg, 48% yield): ¹HNMR (CD₃OD, 500 MHz) δ 0.88 (s, 9H),1.00 (d, J=9.77 Hz, 5H), 1.04 (s, 9H), 1.17 (s, 3H), 1.18-1.20 (m, 1H),1.21 (s, 3H), 1.35 (d, J=2.44 Hz, 1H), 1.40-1.43 (m, 1H), 1.57 (d,J=14.95 Hz, 1H), 1.67 (d, J=14.65 Hz, 1H), 1.85 (dd, J=8.09, 5.34 Hz,1H), 2.15 (d, J=8.24 Hz, 1H), 2.34-2.43 (m, 1H), 2.60 (dd, J=13.73, 7.02Hz, 1H), 2.89-2.93 (m, 1H), 3.92 (s, 3H), 4.13 (dd, J=11.60, 3.97 Hz,1H), 4.38 (s, 1H), 4.43 (d, J=11.90 Hz, 1H), 4.50 (dd, J=9.77, 7.32 Hz,1H), 5.07 (d, J=10.38 Hz, 1H), 5.24 (d, J=17.09 Hz, 1H), 5.75-5.81 (m,1H), 5.84 (s, 1H), 7.09 (dd, J=9.16, 2.44 Hz, 1H), 7.17 (d, J=2.44 Hz,1H), 7.23 (d, J=5.80 Hz, 1H), 7.88 (d, J=5.80 Hz, 1H), 8.10 (d, J=9.16Hz, 1H)); LC-MS (retention time: 1.92 min.), MS m/z 769 (MH⁺).

Example 82 Preparation of Compound 82

Compound 82 was prepared by the same method as Compound 76 with thefollowing modifications:

Modifications: 1-(4-fluorophenyl)-2-methyl-2-propylamine was used as astarting material to give Compound 82 (14.8 mg, 42% yield): ¹HNMR(CD₃OD, 500 MHz) δ 0.88 (s, 9H), 1.00 (d, J=9.46 Hz, 6H), 1.04 (s, 9H),1.17 (s, 3H), 1.21 (s, 3H), 1.32-1.37 (m, 2H), 1.39-1.43 (m, 1H), 1.57(d, J=14.65 Hz, 1H), 1.67 (d, J=14.96 Hz, 1H), 1.82-1.86 (m, 1H), 2.15(t, J=9.46 Hz, 1H), 2.33-2.43 (m, 2H), 2.58-2.62 (dd, J=14.50, 7.78 Hz,1H), 2.89-2.93 (m, 1H), 3.92 (s, 3H), 4.12 (dd, J=11.90, 3.97 Hz, 1H),4.38 (s, 1H), 4.43 (d, J=12.82 Hz, 1H), 4.49-4.52 (m, 1H), 5.24 (d,J=16.48 Hz, 1H), 5.76-5.82 (m, 1H), 5.83-5.85 (m, 1H), 7.09 (dd, J=9.00,2.59 Hz, 1H), 7.17 (d, J=2.14 Hz, 1H), 7.23 (d, J=5.80 Hz, 1H), 7.88 (d,J=5.80 Hz, 1H), 8.10 (d, J=9.16 Hz, 1H)); LC-MS (retention time: 1.40min.), MS m/z 807 (MH⁺).

Example 83 Preparation of Compound 83

Compound 83 was prepared by the same method as Compound 76 with thefollowing modifications:

Modifications: Cumylamine was used as a starting material to giveCompound 83 (64.6 mg, 57% yield): ¹HNMR (CD₃OD, 500 MHz) δ 0.87-0.91 (m,1H), 0.98 (d, J=9.46 Hz, 2H), 1.01 (s, 9H), 1.02-1.05 (m, 1H), 1.17-1.21(m, 3H), 1.29 (s, 2H), 1.40 (dd, J=9.46, 5.19 Hz, 1H), 1.51 (d, J=3.05Hz, 5H), 1.85 (dd, J=8.09, 5.34 Hz, 1H), 2.17 (q, J=8.85 Hz, 1H),2.30-2.33 (m, 1H), 2.58 (dd, J=13.58, 7.48 Hz, 1H), 2.91-2.94 (m, 1H),3.92 (d, J=2.14 Hz, 1H), 3.93 (s, 3H), 4.05 (dd, J=11.60, 3.66 Hz, 1H),4.32 (d, J=9.77 Hz, 1H), 4.51 (dd, J=9.92, 7.17 Hz, 1H), 5.09 (dd,J=11.59, 1.52 Hz, 1H), 5.27 (dd, J=16.71, 1.22 Hz, 1H), 5.74-5.78 (m,1H), 5.82 (s, 1H), 7.05-7.09 (m, 1H), 7.18 (d, J=2.44 Hz, 1H), 7.19 (d,J=7.33 Hz, 1H), 7.22 (d, J=5.80 Hz, 1H), 7.33 (d, J=7.63 Hz, 1H), 7.84(d, J=5.80 Hz, 1H); LC-MS (retention time: 1.76 min.), MS m/z 775 (MH⁺).

Example 84 Preparation of Compound 84

To a solution of the product of step 5 of example 11 (77.0 mg, 0.136mmol), DIEA (70.4 mg, 0.544 mmol) and HATU (77.5 mg, 0.204 mmol) wasadded Boc-MeIle-OH (43.4 mg, 0.177 mmol). After stirring at rt for 14hr, the reaction mixture was washed with 5% aqueous NaHCO₃ (1 mL). Theaqueous layer was extracted with 2×2 mL DCM. The combined organic layerwas washed with 5% aqueous citric acid (2 mL), brine, dried over MgO₄,concentrated and purified by flash column chromatography (SiO₂, 97:3DCM:MeOH) to give Compound 84 (68.4 mg, 69% yield): ¹H NMR (CD₃OD, 500MHz) δ 0.89 (t, J=7.32 Hz, 3H) 0.94 (dd, J=5.95, 4.43 Hz, 3H) 1.07 (d,J=7, 63 Hz, 3H) 1.13 (s, 5H) 1.16-1.20 (m, J=4.88 Hz, 2H) 1.23 (s, 3H)1.28 (m, 1H) 1.34-1.38 (m, 1H) 1.41-1.47 (m, 1H) 1.55-1.60 (m, J=7.63Hz, 1H) 1.87-1.91 (m, 1H) 2.22-2.26 (m, 2H) 2.36-2.38 (m, 1H) 2.56-2.62(m, 1H) 2.81 (d, J=11.30 Hz, 2H) 2.94-2.99 (m, 1H) 3.92 (s, 3H)4.05-4.12 (m, 2H) 4.48-4.57 (m, 2H) 5.12 (d, J=10.07 Hz, 1H) 5.32 (m,1H) 5.75-5.82 (m, 1H) 5.84-5.88 (m, 1H) 7.09-7.13 (m, 1H) 7.16-7.20 (m,1H) 7.23-7.27 (m, 1H) 7.88 (dd, J=5.95, 2.29 Hz, 1H) 8.04 (d, J=9.16 Hz,0.6H) 8.09 (d, J=9.46 Hz, 0.4H); LC-MS (retention time: 1.83 min.), MSm/z 728 (MH⁺).

Example 85 Preparation of Example 85

Compound 85 was prepared by the same method as Compound 84 with thefollowing modifications:

Modifications: Boc-MeVal-OH was used as a starting material to giveCompound 84 (72.1 mg, 74% yield): ¹H NMR (CD₃OD, 500 MHz) δ 0.84 (t,J=5.80 Hz, 3H), 0.96 (d, J=6.41 Hz, 3H), 1.08 (d, J=7.32 Hz, 2H), 1.13(s, 6H), 1.16 (s, 4H), 1.18-1.21 (m, 1H), 1.23-1.29 (m, 1H), 1.44 (dd,J=9.61, 5.34 Hz, 1H), 1.88-1.92 (m, 1H), 2.24 (d, J=10.07 Hz, 1H),2.32-2.39 (m, 2H), 2.58 (dd, J=13.89, 6.26 Hz, 1H), 2.80 (s, 3H),2.93-2.98 (m, 1H), 3.93 (s, 3H), 4.01 (dd, J=11.90, 3.36 Hz, 0.6H), 4.12(dd, J=11.90, 3.66 Hz, 0.4H), 4.16 (d, J=11.29 Hz, 0.6H), 4.38 (d,J=10.99 Hz, 0.4H), 4.45 (d, J=10.68 Hz, 1H), 4.47 (d, J=10.69 Hz, 0.6H),4.53 (dd, J=10.38, 7.02 Hz, 0.4H), 4.58 (dd, J=10.07, 7.02 Hz, 1H), 5.12(d, J=4.28, 0.6H), 5.14 (d, J=4.27 Hz, 0.4H), 5.30 (d, J=7.32 Hz, 0.6H),5.34 (d, J=7.32 Hz, 0.4H), 5.78-5.85 (m, 1H), 5.88 (t, J=3.05 Hz, 0.6H),5.96 (t, J=3.97 Hz, 0.4H), 7.13 (dd, J=9.00, 2.29 Hz, 0.6H), 7.16 (dd,J=9.46, 2.44 Hz, 0.4H), 7.19 (m, 1H), 7.24 (d, J=6.10 Hz, 0.6H), 7.26(d, J=6.10 Hz, 0.4H), 7.88 (d, J=5.80 Hz, 1H), 8.02 (d, J=9.16 Hz, 0.6H)8.05 (d, J=9.16 Hz, 0.4H).

Example 86 Preparation of Compound 86

Compound 86 was prepared by the same method as Compound 84 with thefollowing modifications:

Modifications: Boc-MeLeu-OH was used as a starting material to giveCompound 85 (56.5 mg, 57% yield): ¹H NMR (CD₃OD, 500 MHz) δ 0.94-0.96(m, 6H), 1.04-1.13 (m, 2H), 1.17 (s, 4.5H), 1.18 (s, 4.5H), 1.26-1.31(m, 1H), 1.42 (dd, J=9.46, 5.49 Hz, 1H), 1.46-1.51 (m, 2H), 1.56-1.60(m, 0.5H), 1.69-1.72 (m, 0.5H), 1.75-1.81 (m, 0.5H), 1.90 (q, J=7.50 Hz,1H), 2.27 (dd, J=13.89, 7.78 Hz, 1H), 2.32-2.38 (m, 1H), 2.58 (dd,J=14.80, 7.48 Hz, 1H), 2.75 (s, 3H), 2.95-2.99 (m, 1H), 3.93 (s, 3H),4.03 (d, J=12.21 Hz, 1H), 4.11-15 (m, 0.5H), 4.28 (d, J=12.21 Hz, 1H),4.53 (t, J=8.50 Hz, 0.5H), 4.59 (t, J=8.55 Hz, 0.5H), 4.83-4.87 (m,J=6.41 Hz, 0.5H), 4.96 (m, 0.5H), 5.14 (dd, J=11.14, 4.73 Hz, 1H), 5.32(dd, J=17.70, 6.41 Hz, 1H), 5.75-5.82 (m, 1H), 5.90 (s, 0.5H), 5.92 (s,0.5H), 7.13-7.18 m, 1H), 7.20 (s, 1H), 7.25-7.27 (m, 1H), 7.87 (t,J=4.40 Hz, 1H) 8.05 (d, J=8.85 Hz, 1H).

Example 87 Preparation of Compound 87

Compound 87 was prepared by the same method as Compound 84 with thefollowing modifications:

Modifications: Boc-MeNle-OH was used as a starting material to giveCompound 87 (82.3 mg, 83% yield): ¹H NMR (CD₃OD, 500 MHz) δ 0.90-0.96(q, J=7.63 Hz, 3H) 1.05-1.10 (m, 2H) 1.18 (s, 4.5H) 1.20 (s, 4.5H)1.24-1.30 (m, 3H) 1.31-1.38 (m, 1H) 1.42 (dd, J=9.46, 5.19 Hz, 2H)1.72-1.81 (m, 2H) 1.88-1.92 (m, 1H) 2.22-2.29 (m, 1H) 2.32-2.38 (m, 1H)2.58 (dd, J=13.89, 7.17 Hz, 1H) 2.72 (s, 3H) 2.94-2.99 (m, 1H) 3.93 (s,3H) 4.02 (dd, J=9.77, 4.27 Hz, 1H) 4.12 (dd, J=11.90, 3.35 Hz, 0.5H)4.24 (dd, J=11.90, 0.6 Hz, 0.5H) 4.51-4.60 (m, 1H) 5.14 (d, J=10.38 Hz,1H) 5.33 (dd, J=17.24, 4.73 Hz, 1H) 5.75-5.82 (m, 1H) 5.91 (s, 1H) 7.15(dd, J=14.34, 7.63 Hz, 1H) 7.20 (d, J=2.44 Hz, 1H) 7.25 (d, J=5.80 Hz,1H) 7.87 (t, J=4.37 Hz, 1H) 8.05 (d, J=8.85 Hz, 1H).

Example 88 Preparative of Compound 88

Compound 88 was prepared by the same method as Compound 84 with thefollowing modifications:

Modifications: Boc-N-Me-NVa-OH was used as a starting material to giveCompound 88 (70.5 mg, 73% yield): ¹H NMR (CD₃OD, 500 MHz) δ 0.96 (d,J=6.41 Hz, 3H) 1.06-1.10 (m, 2H) 1.18 (s, 9H) 1.27-1.30 (m, 4H) 1.42(dd, J=9.46, 5.49 Hz, 1H) 1.66-1.80 (m, 2H) 1.88-1.92 (m, 1H) 2.23-2.29(m, 1H) 2.30-2.37 (m, 1H) 2.58 (dd, J=13.58, 7.17 Hz, 1H) 2.73 (s, 3H)2.94-2.98 (m, 1H) 3.93 (s, 3H) 4.00-4.04 (m, 1H) 4.12 (d, J=12.82 Hz,0.5H) 4.25 (d, J=12.21 Hz, 0.5H) 4.51-4.60 (m, 1H) 5.13 (d, J=10.68 Hz,1H) 5.32 (d, J=17.09 Hz, 1H) 5.75-5.81 (m, 1H) 5.90 (s, 1H) 7.13-7.18(m, 1H) 7.20 (d, J=2.14 Hz, 1H) 7.25 (d, J=5.80 Hz, 1H) 7.87 (s, 1H)8.05 (d, J=9.16 Hz, 1H).

Example 89 Preparation of Compound 89

To a solution of the product from step 5 of example 11 (66.0 mg, 0.123mmol), DIEA (63.7 mg, 0.492 mmol) and HATU (70.0, 0.184 mmol) was added2S-tert-butoxycarbonylamino-3-hydroxy-3-methyl-butyric acid (34.0 mg,0.147 mmol). After stirring at rt for 14 hr, the reaction mixture waswashed with 5% aqueous NaHCO₃ (1 mL). The aqueous layer was extractedwith 2×2 mL DCM. The combined organic layer was washed with 5% aqueouscitric acid (2 mL), brine, dried over MgO₄, concentrated and purified byreversed phase prep-HPLC to give Compound 89 (36.1 mg, 41% yield): ¹HNMR (CD₃OD, 500 MHz) δ 1.07 (d, J=7.93 Hz, 2H), 1.18 (s, 1H), 1.20 (s,9H), 1.24-1.27 (m, J=11.60 Hz, 3H), 1.30 (s, 3H), 1.43-1.48 (m, 10H),1.59 (s, 1H), 1.65 (s, 1H), 1.87 (dd, J=8.24, 5.19 Hz, 1H), 2.24 (q,J=9.16 Hz, 1H), 2.33-2.36 (m, 1H), 2.63 (dd, J=12.97, 6.56 Hz, 1H),2.94-2.99 (m, 1H), 3.92 (s, 3H), 3.93 (s, 1H), 4.12 (dd, J=11.60, 3.05Hz, 1H), 4.27-4.31 (m, 1H), 4.54 (t, J=9.77 Hz, 1H), 5.12 (dd, J=10.53,1.37 Hz, 1H), 5.30 (d, J=17.09 Hz, 1H), 5.79-5.83 (m, 1H), 5.85 (s, 1H),7.11 (dd, J=8.55, 1.83 Hz, 1H), 7.18 (d, J=2.24 Hz, 1H), 7.24 (d, J=5.49Hz, 1H), 7.88 (m, 1H), 8.10 (d, J=8.85 Hz, 1H)); LC-MS (retention time:1.637 min.), MS m/z 716 (MH⁺).

Example 91 Preparation of Compound 91

Compound 91 was prepared by the same method as Compound 89 with thefollowing modifications:

Modifications: Boc-L-Thr-OH was used as a starting material to giveCompound 91 (80.5 mg, 66% yield): ¹H NMR (CD₃OD, 500 MHz) δ 0.93 (dd,J=8.24, 2.14 Hz, 2H), 1.08-1.18 (m, 4H), 1.20 (d, J=6.10 Hz, 3H), 1.29(s, 9H), 1.32 (dd, J=9.61, 5.04 Hz, 1H), 1.45 (d, J=4.27 Hz, 1H), 1.84(dd, J=7.63, 5.19 Hz, 1H), 2.15 (q, J=8.85 Hz, 1H), 2.42-2.48 (m, 1H),2.64 (dd, J=14.04, 7.63 Hz, 1H), 2.85-2.89 (m, 1H), 3.92 (s, 3H),4.1-4.14 (m, 2H), 4.30 (d, J=4.88 Hz, 1H), 4.38 (d, J=11.60 Hz, 1H),4.60 (t, J=8.55 Hz, 1H), 5.04 (dd, J=10.22, 1.68 Hz, 1H), 5.80-5.84 (m,2H), 7.11 (d, J=9.16 Hz, 1H), 7.17 (d, J=1.83 Hz, 1H), 7.23 (d, J=5.80Hz, 1H), 7.87 (d, J=5.80 Hz, 1H), 8.10 (d, J=8.85 Hz, 1H); LC-MS(retention time: 1.560 min.), MS m/z 702 (MH⁺).

Example 92 Preparation of Compound 92

Compound 92 was prepared by the same method as Compound 89 with thefollowing modifications:

Modifications: Boc-L-Thr(Me)-OH was used as a starting material to giveCompound 92 (47.1 mg, 69% yield): ¹H NMR (CD₃OD, 500 MHz) δ 0.95 (d,J=4.27 Hz, 2H), 1.11-1.16 (m, 3H), 1.18 (d, J=6.10 Hz, 6H), 1.32 (s,9H), 1.38 (dd, J=9.31, 5.04 Hz, 1H), 1.45 (s, 1H), 1.85 (dd, J=7.78,5.04 Hz, 1H), 2.13 (d, J=9.15 Hz, 1H), 2.46-2.51 (m, 1H), 2.63 (dd,J=14.19, 7.78 Hz, 1H), 2.81-2.91 (m, 1H), 3.68-3.73 (m, 1H), 3.92 (s,4H), 4.14 (d, J=12.21 Hz, 1H), 4.35 (d, J=5.80 Hz, 1H), 4.42 (d, J=11.29Hz, 1H), 4.60 (t, J=8.70 Hz, 1H), 5.05 (d, J=10.68 Hz, 1H), 5.24 (dd,J=16.79, 0.92 Hz, 1H), 5.81-5.85 (m, 2H), 7.11 (dd, J=9.16, 0.92 Hz,1H), 7.17 (s, 1H), 7.24 (d, J=5.80 Hz, 1H), 7.88 (d, J=5.80 Hz, 1H),8.11 (d, J=8.55 Hz, 1H); LC-MS (retention time: 1.660 min.), MS m/z 716(MH⁺).

Example 93 Preparation of Compound 93

Compound 93 was prepared by the same method as Compound 89 with thefollowing modifications:

Modifications: Boc-L-Thr(tBu)—OH was used as a starting material to giveCompound 93 (52.7 mg, 53% yield): ¹H NMR (CD₃OD, 500 MHz) δ 1.09 (dd,J=8.24, 2.44 Hz, 2H), 1.20-1.24 (m, 3H), 1.28 (s, 9H), 1.45 (s, 9H),1.89 (dd, J=7.93, 5.19 Hz, 1H), 2.25 (q, J=8.34 Hz, 1H), 2.32-2.36 (m,1H), 2.59 (dd, J=12.82, 6.41 Hz, 1H), 2.95-3.00 (m, 1H), 3.71 (s, 2H),3.92 (s, 3H), 3.93-3.99 (m, 1H), 4.10 (d, J=7.02 Hz, 1H), 4.15 (d,J=11.90 Hz, 1H), 4.22 (dd, J=6.10, 2.44 Hz, 2H), 4.40 (d, J=11.60 Hz,1H), 4.54 (dd, J=10.01, 6.71 Hz, 1H), 5.13 (d, J=10.38 Hz, 1H), 5.31 (d,J=17.09 Hz, 1H), 5.76-5.83 (m, 1H), 5.87 (s, 1H), 6.06 (d, J=9.46 Hz,1H), 6.36 (d, J=7.02 Hz, 1H), 7.11 (d, J=8.85 Hz, 1H), 7.18 (s, 1H),7.24 (d, J=5.49 Hz, 1H), 7.88 (m, 1H), 8.08 (d, J=9.16 Hz, 1H).

Example 94 Preparation of Compound 94

Compound 94 was prepared by the same method as Compound 89 with thefollowing modifications:

Modifications: Boc-(2S,3S)-2-amino-3-methoxybutanoic acid was used as astarting material to give Compound 94 (150.2 mg, 80% yield): ¹H NMR(CD₃OD, 500 MHz) δ 1.04-1.13 (m, 3H), 1.17 (d, J=6.10 Hz, 3H), 1.20-1.24(m, 2H), 1.27 (s, 9H), 1.44-1.48 (m, 2H), 1.86 (dd, J=7.93, 5.49 Hz,1H), 2.24 (q, J=8.65 Hz, 1H), 2.34-2.37 (m, 1H), 2.61 (dd, J=14.19, 7.17Hz, 1H), 2.94-2.99 (m, 1H), 3.66 (m, 1H), 3.92 (s, 3H), 4.13 (dd,J=12.36, 3.81 Hz, 1H), 4.37 (dd, J=22.58, 10.99 Hz, 2H), 4.54 (dd,J=10.38, 7.63 Hz, 1H), 5.12 (d, J=10.68 Hz, 1H), 5.31 (d, J=17.40 Hz,1H), 5.77-5.82 (m, 1H), 5.85 (s, 1H), 7.12 (d, J=9.16 Hz, 1H), 7.18 (s,1H), 7.25 (d, J=6.10 Hz, 1H), 7.89 (d, J=5.80 Hz, 1H), 8.10 (d, J=8.85Hz, 1H); LC-MS (retention time: 1.673 min.), MS m/z 716 (MH⁺).

Example 95 Preparation of Compound 95

Step 1

To a mixture of H-allo-THr-OH (5.0 g, 41.98 mmol) and DIEA (10.9 g,83.96 mmol) in DCM (150 mL) was added di-tert-butyl dicarbonate (13.7 g,62.97 mmol). After stirring at rt for 14 h, the reaction mixture waswashed with 3×100 mL DCM.

The combined organic layer was dried over MgSO₄ and concentrated. LC/MSindicated most product stayed in the H₂O layer. Thus the water layer wasconcentrated. The product was purified by a flash column chromatography(SiO2, 90:10 DCM:MeOH) to give Boc-allo-THr-OH; LC-MS (retention time:0.727 min.), MS m/z 242 (MNa⁺).

Step 2

To a solution of the product from step 5 of example 11 (100.0 mg, 0.174mmol), DIEA (67.6 mg, 0.522 mmol) and HATU (106.0 mg, 0.278 mmol) wasadded the product from step 1 above (57.3 mg, 0.262 mmol). Afterstirring at rt for 3 hr, the reaction mixture was washed with 5% aqueousNaHCO₃ (1 mL). The aqueous layer was extracted with 2×2 mL DCM. Thecombined organic layer was washed with 5% aqueous citric acid (2 mL),brine, dried over MgO₄, concentrated and purified by reversed phaseprep-HPLC to give Compound 95 (39.1 mg, 32% yield): ¹H NMR (CD₃OD, 500MHz) δ 1.02 (d, J=8.55 Hz, 1H), 1.18-1.23 (m, 3H), 1.25 (s, 9H), 1.38(dd, J=9.15, 6.30 Hz 1H), 1.84 (dd, J=7.93, 5.19 Hz, 1H), 2.19-2.24 (m,1H), 2.38-2.43 (m, 1H), 2.65 (dd, J=14.19, 6.87 Hz, 1H), 2.92-2.96 (m,1H), 3.92 (s, 3H), 3.93 (s, 2H), 4.16-4.19 (m, 1H), 4.23 (d, J=8.24 Hz,1H), 4.44 (d, J=12.21 Hz, 1H), 4.57-5.81 (m, 1H), 5.09 (d, J=10.68 Hz,1H), 5.29 (d, J=17.09 Hz, 1H), 5.75-5.81 (m, 1H), 5.83-5.85 (m, 1H),7.11 (d, J=10.38 Hz, 1H), 7.18 (d, J=1.83 Hz, 1H), 7.24 (d, J=6.41 Hz,1H), 7.88 (d, J=5.80 Hz, 1H), 8.11 (d, J=9.16 Hz, 1H); LC-MS (retentiontime: 1.583 min.), MS m/z 702 (MH⁺).

Example 96 Preparation of Compound 96

Compound 96 was prepared by the same method as Compound 95 with thefollowing modifications:

Modifications:

Step 1

(2S,3S)-2-Amino-3-ethoxybutanoic acid hydrochloride was used as astarting material in step 1 to give Boc-(2S,3S)-2-Amino-3-ethoxybutanoicacid; LC-MS (retention time: 1.067 min.), MS m/z 270 (M+Na⁺).

Step 2

The product from step 1 was then coupled the same way with the productfrom step 5 of example 11 to give Compound 96 (55.3 mg, 44% yield): ¹HNMR (CD₃OD, 500 MHz) δ 0.94 (t, J=6.87 Hz, 1H), 0.97-1.03 (m, 2H),1.08-1.11 (m, 2H), 1.13-1.15 (m, 2H), 1.17 (d, J=6.10 Hz, 6H), 1.29 (s,9H), 1.41-1.45 (m, 3H), 1.85 (dd, J=7.48, 5.34 Hz, 1H), 2.12-2.19 (m,1H), 2.43-2.49 (m, 1H), 2.60 (dd, J=13.73, 6.80 Hz, 1H), 2.89-2.93 (m,1H), 3.50-3.57 (m, 2H), 3.73-3.78 (m, 1H), 3.92 (s, 3H), 4.18 (d, J=8.85Hz, 1H), 4.35 (d, J=12.21 Hz, 1H), 4.39 (d, J=8.55 Hz, 1H), 4.53 (t,J=7.78 Hz, 1H), 5.07 (d, J=9.16 Hz, 1H), 5.25 (d, J=18.01 Hz, 1H), 5.82(t, J=9.85 Hz, 1H), 5.88 (t, J=9.80 Hz, 1H), 7.11 (d, J=5.19 Hz, 1H),7.18 (d, J=2.14 Hz, 1H), 7.24 (d, J=5.49 Hz, 1H), 7.88 (d, J=6.10 Hz,1H), 8.10 (d, J=8.85 Hz, 1H); LC-MS (retention time: 1.743 min.), MS m/z730 (MH⁺).

Example 97 Preparation of Compound 97

Compound 97 was prepared by the same method as Compound 95 with thefollowing modifications:

Modifications:

Step 1

H-allo-Thr(t-Bu)—OH was used as a starting material in step 1 to giveBoc-(2S,3S)-2-Amino-3-ethoxybutanoic acid; LC-MS (retention time: 1.363min.), MS m/z 298 (M+Na⁺).

Step 2

The product from step 1 was then coupled the same way with the productfrom step 5 of example 11 to give Compound 97 (48.2 mg, 37% yield):LC-MS (retention time: 1.820 min.), MS m/z 758 (MH⁺).

Example 99 Preparation of Compound 99

Compound 99 was prepared by the same method as step 1 of Example 55 withthe following modifications:

Modifications: Compound 84 was used as a starting material to giveCompound 99 (60.3 mg, 98% yield): ¹H NMR (CD₃OD, 500 MHz) δ 1.00 (q,J=7.12 Hz, 3H) 1.10-113 (m, 5H) 1.20-1.31 (m, 3H) 1.41 (dd, J=9.46, 5.49Hz, 1H) 1.61-1.68 (m, 1H) 1.92 (dd, J=8.24, 5.49 Hz, 1H) 2.04-2.09 (m,1H) 2.28 (q, J=8.55 Hz, 1H) 2.34-2.39 (m, 1H) 2.57 (s, 3H) 2.64-2.70 (m,1H) 2.94-2.97 (m, 1H) 3.93 (s, 3H) 4.07-4.14 (dd, J=12.05, 3.81 Hz, 1H)4.13 (d, J=6.10 Hz, 1H) 4.18 (d, J=5.80 Hz, 1H) 4.25 (d, J=12.21 Hz, 1H)4.66-4.73 (m, 1H) 5.15 (d, J=10.68 Hz, 1H) 5.32 (d, J=17.09 Hz, 1H)5.70-5.79 (m, 1H) 5.92 (t, J=3.66 Hz, 0.4H) 5.95 (t, J=3.66 Hz, 0.6H)7.17 (dd, J=9.16, 2.44 Hz, 1H) 7.22 (d, J=2.14 Hz, 1H) 7.28 (dd, J=5.80,3.36 Hz, 1H) 7.91 (dd, J=5.80, 4.27 Hz, 1H) 8.03 (d, J=8.85 Hz, 0.6H)8.07 (d, J=9.16 Hz, 0.4H); LC-MS (retention time: 1.33 min.), MS m/z6.28 (MH⁺).

Example 100 Preparation of Compound 100

To a solution of Compound 94 (0.600 g, 0.838 mmol) in DCE (3 mL) wasadded TFA (3 mL). After stirring at rt for 15 min, the reaction mixturewas concentrated. The resulting viscous oil was re-dissolved in DCE (5mL) and reconcentrated. It was then redissolved in DCM (2 mL) andtreated with a solution of 1N HCl in Et₂O (10 mL). The resultingsuspension was chilled at 0° C., vacuum filtrated, washed with Et₂O anddried in vacuum oven to give the product as a bis-hydrochloride salt asa white solid (527.1 g, 91% yield): ¹H NMR (CD₃OD, 500 MHz) δ 1.08-1.15(m, 2H), 1.21 (d, J=6.71 Hz, 4H), 1.28-1.33 (m, 1H), 1.41 (dd, J=9.46,5.49 Hz, 1H), 1.91 (dd, J=8.24, 5.49 Hz, 1H), 2.28 (q, J=8.65 Hz, 1H),2.34-2.37 (m, 1H), 2.68 (dd, J=13.12, 7.02 Hz, 1H), 2.81 (s, 3H),2.93-2.98 (m, 1H), 3.45 (s, 3H), 3.94 (s, 3H), 3.96-4.00 (m, 1H), 4.16(dd, J=11.90, 3.66 Hz, 1H), 4.27 (d, J=11.60 Hz, 1H), 4.59 (d, J=4.58Hz, 1H), 4.69 (dd, J=10.07, 7.02 Hz, 1H), 5.14 (dd, J=10.53, 1.37 Hz,1H), 5.32 (d, J=17.09 Hz, 1H), 5.70-5.77 (m, 1H), 5.94 (t, J=3.66 Hz,1H), 7.19 (d, J=9.16 Hz, 1H), 7.24 (s, 1H), 7.32 (s, 1H), 7.91 (d,J=5.80 Hz, 1H), 8.09 (d, J=9.16 Hz, 1H); LC-MS (retention time: 1.213min.), MS m/z 616 (MH⁺).

Example 101 Preparation of Compound 101

To a solution mixture of Compound 100 (80 mg, 0.116 mmol) and DIEA (31.5mg, 0.244 mmol) in THF (2 mL) was added N,N′-dissucinimidyl carbonate(44.6 mg, 0.174 mmol). The resulting suspension was irradiated in amicrowave to 80° C. for min. Then it was treated with tert-amylamine(84.8 mg, 1.16 mmol). After stirring at rt 15 min, the reaction wasconcentrated and purified by reversed phase prep-HPLC to give Compound101 (65.1 mg, 79%): ¹H NMR (CD₃OD, 500 MHz) δ 1.03-1.08 (m, 3H), 1.16(d, J=6.41 Hz, 3H), 1.19 (s, 9H), 1.21-1.25 (m, 2H), 1.27 (d, J=6.10 Hz,1H), 1.45 (dd, J=9.46, 5.19 Hz, 1H), 1.85 (dd, J=8.24, 5.19 Hz, 1H),2.23 (q, J=9.46 Hz, 1H), 2.34-2.41. (m, 1H), 2.61 (dd, J=14.34, 7.32 Hz,1H), 2.94-2.97 (m, 1H), 3.58-3.63 (m, 1H), 3.92 (s, 3H), 4.15 (dd,J=12.05, 3.81 Hz, 1H), 4.39 (d, J=11.60 Hz, 1H), 4.55 (dd, J=9.92, 7.48Hz, 1H), 5.11 (d, J=10.68 Hz, 1H), 5.29 (d, J=17.40 Hz, 1H), 5.77-5.83(m, 1H), 5.86 (s, 1H), 7.12 (dd, J=9.00, 2.29 Hz, 1H), 7.18 (d, J=2.14Hz, 1H), 7.24 (d, J=5.80 Hz, 1H), 7.88 (d, J=5.80 Hz, 1H), 8.10 (d,J=9.16 Hz, 1H); LC-MS (retention time: 1.617 min.), MS m/z 715 (MH⁺).

Example 102 Preparation of Compound 102

Compound 102 was prepared by the same method as Compound 101 with thefollowing modifications:

Modifications: tert-amylamine was used as a starting material to giveCompound 102 (62.5 mg, 74% yield): ¹H NMR (CD₃OD, 500 MHz) δ 0.77 (t,J=7.48 Hz, 2H), 0.84 (t, J=7.48 Hz, 1H), 1.04-1.08 (m, 2H), 1.13 (d,J=1.22 Hz, 9H), 1.16 (d, J=6.41 Hz, 3H), 1.21 (s, 1H), 1.22-1.28 (m,2H), 1.44 (dd, J=9.46, 5.19 Hz, 1H), 1.52-1.57 (m, 1H), 1.58-1.62 (m,1H), 1.85 (dd, J=7.93, 5.19 Hz, 1H), 2.21-2.25 (m, 1H), 2.34-2.39 (m,1H), 2.61 (dd, J=13.58, 7.17 Hz, 1H), 2.93-2.98 (m, 1H), 3.59-3.64 (m,1H), 4.15 (dd, J=11.75, 3.81 Hz, 1H), 4.38 (d, J=12.51 Hz, 1H), 4.50 (d,J=7.63 Hz, 1H), 4.55 (dd, J=9.92, 7.78 Hz, 1H), 5.11 (d, J=9.77 Hz, 1H),5.29 (d, J=16.79 Hz, 1H), 5.77-5.83 (m, 1H), 5.86 (t, J=4.73 Hz, 1H),7.12 (dd, J=8.85, 2.44 Hz, 1H), 7.18 (d, J=2.44 Hz, 1H), 7.24 (d, J=6.10Hz, 1H), 7.88 (d, J=5.80 Hz, 1H), 8.10 (d, J=9.16 Hz, 1H); LC-MS(retention time: 1.690 min.), MS m/z 729 (MH⁺).

Example 103 Preparation of Compound 103

Compound 103 was prepared by the same method as Compound 101 with thefollowing modifications:

Modifications: cyclopentylamine was used as a starting material to giveCompound 103 (56.4 mg, 67% yield): ¹H NMR (CD₃OD, 500 MHz) δ 1.01-1.08(m, 2H), 1.07 (d, J=6.10 Hz, 1H), 1.16 (d, J=6.10 Hz, 3H), 1.21-1.25 (m,3H), 1.30-1.33 (m, 1H), 1.44 (dd, J=9.77, 5.19 Hz, 1H), 1.51-1.56 (m,2H), 1.60-1.65 (m, 2H), 1.71-1.75 (m, 1H), 1.80-1.84 (m, 1H), 1.86 (dd,J=8.09, 5.34 Hz, 1H), 2.20-2.25 (m, 1H), 2.37-2.41 (m, 1H), 2.61 (dd,J=14.04, 7.32 Hz, 1H), 2.93-2.98 (m, 1H), 3.60-3.65 (m, 1H), 3.75-3.80(m, 1H), 3.92 (s, 3H), 4.17 (dd, J=12.05, 3.81 Hz, 1H), 4.37 (d, J=11.90Hz, 1H), 4.55-4.59 (m, 2H), 5.10 (d, J=11.60 Hz, 1H), 5.29 (d, J=16.48Hz, 1H), 5.78-5.83 (m, 1H), 5.85 (d, J=2.44 Hz, 1H), 7.13 (dd, J=9.16,2.44 Hz, 1H), 7.18 (d, J=2.44 Hz, 1H), 7.24 (d, J=5.80 Hz, 1H), 7.88 (d,J=5.80 Hz, 1H), 8.09 (d, J=9.16 Hz, 1H); LC-MS (retention time: 1.607min.), MS m/z 727 (MH⁺).

Example 104 Preparation of Compound 104

To a solution mixture of Compound 100 (80 mg, 0.116 mmol) and DIEA (31.5mg, 0.244 mmol) in THF (2 mL) was added N,N′-dissucinimidyl carbonate(44.6 mg, 0.174 mmol). The resulting suspension was irradiated in amicrowave to 80° C. for min. Then was added a slurry solution of sodiumcyclopentoxide which was prepared by treating a 0° C. solution ofcyclopentanol (110 mg, 1.28 mmol) in THF (1 mL) with NaH (60% in oil,46.4 mg, 1.16 mmol) for 15 min at rt. After stirring at rt min, thereaction was quenched with saturated aqueous ammonium chloride (1 mL)and extracted with EtOAc (5 mL). The organic layer was then passedthrough a celite hydromatrix column, concentrated and purified byreversed phase prep-HPLC to give Compound 104 (38.2 mg, 45%): ¹H NMR(CD₃OD, 500 MHz) δ 1.03-1.09 (m, 3H), 1.16 (d, J=6.10 Hz, 3H), 1.20-1.25(m, 1H), 1.25-1.30 (m, J=10.22, 5.34 Hz, 1H), 1.40-1.45 (m, J=10.83,3.81 Hz, 1H), 1.46 (dd, J=9.61, 5.34 Hz, 1H), 1.58-1.63 (m, 3H),1.70-1.75 (m, 2H), 1.86 (dd, J=7.63, 5.49 Hz, 1H), 2.22-2.26 (m, 1H),2.34-2.39 (m, 1H), 2.59-2.64 (m, 1H), 2.94-2.98 (m, 1H), 3.67 (dd,J=7.78, 6.56 Hz, 1H), 3.92 (s, 3H), 4.13 (dd, J=10.83, 4.12 Hz, 1H),4.37-4.42 (m, 1H), 4.56 (dd, J=10.07, 7.32 Hz, 1H), 4.71-4.76 (m, 1H),5.12 (d, J=10.68 Hz, 1H), 5.31 (d, J=16.79 Hz, 1H), 5.80 (m, 1H), 5.85(s, 1H), 7.13 (d, J=10.68 Hz, 1H), 7.19 (d, J=1.83 Hz, 1H), 7.25 (d,J=6.10 Hz, 1H), 7.89 (d, J=5.80 Hz, 1H), 8.09 (d, J=9.16 Hz, 1H), LC-MS(retention time: 1.697 min.), MS m/z 728 (MH⁺).

Example 105 Preparation of Compound 105

To a solution mixture of Compound 100 (80.0 mg, 0.116 mmol) and DIEA(31.5 mg, 0.244 mmol) in DCM (2 mL) was added di-tert-amyl dicarbonate(57.1 mg, 0.232 mmol). After stirring at rt for 14 h, solvent wasremoved and product was purified by reversed phase prep-HPLC to giveCompound 105 (62.5 mg, 74% yield):

¹H NMR (CD₃OD, 500 MHz) δ 0.79 (t, J=7.48 Hz, 3H), 1.04-1.08 (m, 3H),1.17 (d, J=6.10 Hz, 3H), 1.19-1.23 (s, 3H), 1.24 (s, 3H), 1.39-1.43 (m,1H), 1.46 (dd, J=9.61, 5.34 Hz, 1H), 1.60-1.65 (m, 2H), 1.86 (dd,J=7.93, 5.49 Hz, 1H), 2.22 (q, J=8.85 Hz, 1H), 2.35-2.40 (m, 1H), 2.61(dd, J=14.04, 7.15 Hz, 1H), 2.94-3.00 (m, 1H), 3.64-4.00 (m, 1H), 3.92(s, 4H), 4.14 (dd, J=11.90, 3.05 Hz, 1H), 4.35 (d, J=7.93 Hz, 1H), 4.40(d, J=11.90 Hz, 1H), 4.55 (dd, J=9.31, 7.78 Hz, 1H), 5.11 (d, J=10.68Hz, 1H), 5.30 (d, J=16.79 Hz, 1H), 5.79-5.83 (m, 1H), 5.85 (s, 1H), 7.12(d, J=9.16 Hz, 1H), 7.18 (s, 1H), 7.25 (d, J=5.80 Hz, 1H), 7.86-7.90 (m,1H), 8.09 (d, J=9.16 Hz, 1H); LC-MS (retention time: 1.740 min.), MS m/z730 (MH⁺).

Example 106 Preparation of Compound 106

Compound 106 was prepared by the same method as Compound 105 with thefollowing modifications:

Modifications: Carbonic acid pyridin-2-yl ester2,2,2-trifluoro-1,1-dimethyl-ethyl ester was used as a starting materialto give Compound 106 (58.1 mg, 65% yield): ¹H NMR (CD₃OD, 500 MHz) δ1.04-1.08 (m, 3H), 1.17 (d, J=6.10 Hz, 3H), 1.19-1.23 (m, 1H), 1.23-1.27(m, 1H), 1.28 (s, 3H), 1.46 (dd, J=9.46, 5.19 Hz, 2H), 1.49 (s, 2H),1.86 (dd, J=8.09, 5.34 Hz, 1H), 2.21 (q, J=8.85 Hz, 1H), 2.36-2.40 (m,1H), 2.62 (dd, J=13.74, 7.32 Hz, 1H), 2.93-2.98 (m, 1H), 3.65-3.70 (m,1H), 3.92 (s, 3H), 4.12 (dd, J=11.90, 3.66 Hz, 1H), 4.31 (d, J=8.24 Hz,1H), 4.42 (d, J=11.90 Hz, 1H), 4.57 (dd, J=10.07, 7.32 Hz, 1H), 5.11 (d,J=10.38 Hz, 1H), 5.30 (d, J=16.79 Hz, 1H), 5.78-5.83 (m, 1H), 5.84 (s,1H), 7.12 (dd, J=9.00, 2.29 Hz, 1H), 7.19 (d, J=2.14 Hz, 1H), 7.25 (d,J=5.80 Hz, 1H), 7.89 (d, J=6.10 Hz, 1H), 8.09 (d, J=8.85 Hz, 1H); LC-MS(retention time: 1.770 min.), MS m/z 770 (MH⁺).

Example 107 Preparation of Compound 107

To a solution of the product from step 3 of Example 25 (100.0 mg, 0.116mmol), DIEA (62.5 mg, 0.483 mmol) and HATU (92.0 mg, 0.242 mmol) wasadded Boc-allo-Thr-OH (43.5 mg, 0.177 mmol). After stirring at rt for 3hr, the reaction mixture was washed with 5% aqueous NaHCO₃ (1 mL). Theaqueous layer was extracted with 2×2 mL DCM. The combined organic layerwas washed with 5% aqueous citric acid (1 mL), brine, dried over MgO₄,concentrated and purified by reversed phase prep-HPLC to give Compound107 (62.5 mg, 52% yield): ¹HNMR (CD₃OD, 500 MHz) δ0.8-1.02 (m, 1H),1.04-1.08 (m, 2H), 1.23-1.27 (m, 12H), 1.42 (dd, J=9.46, 5.19 Hz, 1H),1.86 (t, J=6.26 Hz, 1H), 2.23-2.27 (m, 1H), 2.46-2.50 (m, 1H), 2.76 (dd,J=14.04, 6.71 Hz, 1H), 2.95-2.99 (m, 1H), 3.94-3.98 (m, 1H), 4.28 (d,J=7.32 Hz, 2H), 4.52 (d, J=12.51 Hz, 1H), 4.63 (t, J=9.00 Hz, 1H), 5.12(d, J=10.07 Hz, 1H), 5.31 (d, J=16.79 Hz, 1H), 5.77-5.83 (m, 1H), 6.09(s, 1H), 7.36-7.41 (m, 1H), 7.47 (t, J=7.17 Hz, 3H), 7.52 (d, J=7.63 Hz,1H), 7.70 (t, J=7.17 Hz, 1H), 7.85 (s, 1H), 7.88 (d, J=8.24 Hz, 1H),8.17 (d, J=7.93 Hz, 2H), 8.22 (d, J=7.63 Hz, 1H); LC-MS (retention time:1.937 min.), MS m/z 748 (MH⁺).

Example 108 Preparation of Compound 108

Compound 108 was prepared by the same method as Compound 107 with thefollowing modifications:

Modifications: Boc-(2S,3S)-Amino-3-methoxybutanoic acid was used as astarting material to give Compound 108 (75.1 mg, 51% yield): ¹H NMR(CD₃OD, 500 MHz) δ 0.80-1.02 (m, 4H), 1.18 (d, J=6.10 Hz, 3H), 1.28 (s,9H), 1.44 (dd, J=9.77, 1.30 Hz, 1H), 1.45-1.50 (m, 1H), 1.85-1.90 (m,1H), 2.14-2.18 (m, 1H), 2.55-2.59 (m, 1H), 2.72-2.76 (m, 1H), 2.91-2.95(m, 1H), 3.34 (s, 3H), 3.65-3.69 (m, 1H), 4.32 (d, J=10.68 Hz, 1H), 4.40(d, J=7.93 Hz, 1H), 4.46 (d, J=13.12 Hz, 1H), 4.60 (t, J=8.24 Hz, 1H),5.07 (d, J=9.46 Hz, 1H), 5.26 (d, J=17.40 Hz, 1H), 5.82-5.86 (m, 1H),6.08 (s, 1H), 7.38 (dd, J=7.32, 6.10 Hz, 1H), 7.47 (t, J=7.02 Hz, 3H),7.51 (d, J=5.80 Hz, 1H), 7.69 (t, J=6.56 Hz, 1H), 7.69 (t, J=6.56 Hz,1H), 7.85 (s, 1H), 7.88 (d, J=7.63 Hz, 1H), 8.18 (d, J=8.24 Hz, 3H),8.21 (d, J=9.16 Hz, 1H); LC-MS (retention time: 1.973 min.), MS m/z 762(MH⁺).

Example 109 Preparation of Compound 109

Compound 109 was prepared by the same method as Compound 107 with thefollowing modifications:

Modifications: Boc-(2S,3S)-Amino-3-ethoxybutanoic acid was used as astarting material to give Compound 109 (57.2 mg, 47% yield): ¹H NMR(CD₃OD, 500 MHz) δ 1.02-1.08 (m, 4H), 1.17 (d, J=6.10 Hz, 6H), 1.19 (s,1H), 1.19-1.24 (m, 1H), 1.23-1.27 (m, J=3.97 Hz, 1H), 1.30 (s, 9H), 1.44(dd, J=9.77, 5.50 Hz, 1H), 1.46 (s, 1H), 1.88 (dd, J=7.78, 5.95 Hz, 1H),2.20-1.25 (m, 1H), 2.49-2.54 (m, 1H), 2.69-2.73 (m, 1H), 2.93-2.97 (m,1H), 3.53-3.57 (m, 2H), 3.75-3.80 (m, 1H), 4.34 (dd, J=11.75, 3.20 Hz,1H), 4.42 (t, J=8.30 Hz, 2H), 4.57 (t, J=8.09 Hz, 1H), 5.11 (d, J=10.38Hz, 1H), 5.29 (d, J=17.40 Hz, 1H), 5.78-5.83 (m, 1H), 6.09 (s, 1H), 7.38(t, J=7.32 Hz, 1H), 7.47 (t, J=7.63 Hz, 2H), 7.52 (d, J=7.02 Hz, 1H),7.70 (t, J=7.93 Hz, 1H), 7.86 (s, 1H), 7.88 (d, J=7.93 Hz, 1H), 8.18 (d,J=7.32 Hz, 2H), 8.22 (d, J=8.24 Hz, 1H); LC-MS (retention time: 2.030min.), MS m/z 776 (MH⁺).

Example 110 Preparation of Compound 110

Compound 110 was prepared by the same method as Compound 89 with thefollowing modifications:

Modifications: Boc-L-Thr(Bn)—OH was used as a starting material to giveCompound 110 (49.8 mg, 48% yield)); LC-MS (retention time: 1.857 min.),MS m/z 792 (MH⁺).

Section D Example 120 Preparation of Compound 120

Step 1:

A solution of Compound 23 (see Example 23) (1.50 g, 2.19 mmol) in DCM(50 mL) and trifluoroacetic acid (50 mL) was stirred for 3 h at rt. Themixture was concentrated in vacuo to a viscous residue, and was thendissolved in 1,2-dichloroethane and again concentrated in vacuo to givethe desired bis-trifluoroacetic acid salt product as an off-white glassysolid (quantitative). The material was used directly in the next stepwithout purification.

Step 2:

To a solution of the product from Example 120, Step 1 (118 mg, 0.146mmol) in 1,2-dichloroethane (3 mL) was added p-tolyl chloroformate (32.4mg, 0.190 mmol) and N,N-diisopropylethylamine (94.5 mg, 0.731 mmol). Themixture was agitated at rt for 72 h. The reaction mixture was washedwith pH=4 buffer solution (3×3 mL), and the washes were back-extractedwith 1,2-dichloroethane (3 mL). The organic phases were combined andconcentrated in vacuo. The crude product was then dissolved in MeOH andpurified by reverse phase preparative HPLC to give the title compound(Compound 120) as a yellow glassy solid (64.2 mg, 61.1% yield):

¹H NMR (CD₃OD) δ 1.06-1.10 (m, 3H), 1.12 (s, 9H), 1.24-1.28 (m, 2H),1.44 (dd, J=9.31, 5.34 Hz, 1H), 1.89 (dd, J=7.93, 5.49 Hz, 1H),2.21-2.28 (m, 2H), 2.31 (s, 3H), 2.62-2.66 (m, 1H), 2.93-2.99 (m, 1H),4.12 (dd, J=11.90, 3.66 Hz, 1H), 4.42 (d, J=11.60 Hz, 1H), 4.57 (dd,J=10.22, 7.17 Hz, 1H), 5.13 (d, J=10.38 Hz, 1H), 5.30 (d, J=17.09 Hz,1H), 5.76 (ddd, J=17.09, 9.77, 9.46 Hz, 1H), 5.87 (s, 1H), 6.79 (d,J=8.24 Hz, 2H), 7.07 (d, J=8.24 Hz, 2H), 7.30 (d, J=6.10 Hz, 1H), 7.40(t, J=7.63 Hz, 1H), 7.68 (t, J=7.63 Hz, 1H), 7.79 (d, J=8.24 Hz, 1H),7.93 (d, J=5.80 Hz, 1H), 8.17 (d, J=8.24 Hz, 1H); MS m/z 718 (MH⁺).

Example 121 Preparation of Compound 121

Compound 121 was prepared by following Scheme 1 of Example 120 exceptthat phenyl chloroformate was used in place of p-tolyl chloroformate instep 2.

Step 2:

Modifications: 30 mg (0.19 mmol) phenyl chloroformate used, 89.0 mgproduct obtained as a yellow glassy solid (50% yield): MS m/z 704 (MH⁺).

Example 122 Preparation of Compound 122

Compound 122 was prepared by following Scheme 1 of Example 120 exceptthat 4-fluorophenyl chloroformate was used in place of p-tolylchloroformate in step 2.

Step 2:

Modifications: 33 mg (0.19 mmol) 4-fluorophenyl chloroformate used, 83.1mg product obtained as a sticky yellow oil (78.8% yield): MS m/z 722(MH⁺).

Example 123 Preparation of Compound 123

Compound 123 was prepared by following Scheme 1 of Example 120 exceptthat 4-methoxyphenyl chloroformate was used in place of p-tolylchloroformate in step 2.

Step 2:

Modifications: 35 mg (0.19 mmol) 4-methoxyphenyl chloroformate used,70.2 mg product obtained as a yellow glassy solid (65.4% yield): ¹H NMR(CD₃OD) δ 1.06-1.10 (m, 3H), 1.11 (s, 9H), 1.24-1.28 (m, 2H), 1.44 (dd,J=9.46, 5.49 Hz, 1H), 1.89 (dd, J=7.93, 5.49 Hz, 1H), 2.24 (q, J=8.85Hz, 1H), 2.31 (ddd, J=13.81, 10.30, 3.97 Hz, 1H), 2.62-2.66 (m, 1H),2.94-2.98 (m, 1H), 3.77 (s, 3H), 4.12 (dd, J=11.60, 3.66 Hz, 1H), 4.42(d, J=11.60 Hz, 1H), 4.57 (dd, J=10.07, 7.32 Hz, 1H), 5.13 (d, J=10.68Hz, 1H), 5.30 (d, J=16.79 Hz, 1H), 5.72-5.80 (m, 1H), 5.87 (s, 1H), 6.80(d, J=2.44 Hz, 4H), 7.30 (d, J=5.80 Hz, 1H), 7.42 (t, J=7.48 Hz, 1H),7.69 (t, J=7.63 Hz, 1H), 7.80 (d, J=7.93 Hz, 1H), 7.93 (d, J=5.80 Hz,1H), 8.18 (d, J=8.24 Hz, 1H); MS m/z 734 (MH⁺).

Example 124 Preparation of Compound 124

Compound 124 was prepared by following Scheme 1 of Example 120 exceptthat chloroformic acid 2-methoxyethyl ester was used in place of p-tolylchloroformate in step 2.

Step 2:

Modifications: 26 mg (0.19 mmol) chloroformic acid 2-methoxyethyl esterused, 87.4 mg product obtained as a sticky yellow oil (87.2% yield): ¹HNMR (CD₃OD) δ 0.96-1.02 (m, 3H), 1.05 (s, 9H), 1.16-1.18 (m, 2H), 1.40(dd, J=9.46, 5.19 Hz, 1H), 1.85 (dd, J=7.93, 5.19 Hz, 1H), 2.15 (q,J=8.75 Hz, 1H), 2.40 (ddd, J=13.89, 10.07, 4.12 Hz, 1H), 2.65 (dd,J=13.58, 7.17 Hz, 1H), 2.90 (ddd, J=12.89, 8.16, 4.88 Hz, 1H), 3.27 (s,3H), 3.36-3.44 (m, 2H), 3.81-3.84 (m, 1H), 3.92-3.96 (m, 1H), 4.12 (dd,J=11.60, 3.36 Hz, 1H), 4.44 (d, J=11.60 Hz, 1H), 4.57 (dd, J=9.46, 7.93Hz, 1H), 5.07 (d, J=10.38 Hz, 1H), 5.25 (d, J=17.09 Hz, 1H), 5.80 (ddd,J=17.32, 9.77, 9.54 Hz, 1H), 5.87 (s, 1H), 7.32 (d, J=5.80 Hz, 1H), 7.55(t, J=7.32 Hz, 1H), 7.70 (t, J=7.48 Hz, 1H), 7.80 (d, J=7.93 Hz, 1H),7.96 (d, J=5.80 Hz, 1H), 8.19 (d, J=8.24 Hz, 1H); MS m/z 686 (MH⁺).

Example 125 Preparation of Compound 125

Compound 125 was prepared by following Scheme 1 of Example 120 exceptthat neopentyl chloroformate was used in place of p-tolyl chloroformatein step 2.

Step 2:

Modifications: 29 mg (0.19 mmol) neopentyl chloroformate used, 57.4 mgproduct obtained as a yellow glassy solid (56.2% yield): ¹H NMR (CD₃OD)δ 0.83 (s, 9H), 1.05 (d, J=2.44 Hz, 9H), 1.07-1.09 (m, 2H), 1.23-1.27(m, 2H), 1.43-1.46 (m, 1H), 1.87-1.90 (m, 1H), 2.21-2.25 (m, 1H),2.29-2.33 (m, 1H), 2.61-2.65 (m, 1H), 2.92-2.96 (m, 1H), 3.42 (d,J=10.07 Hz, 1H), 3.56 (d, J=10.07 Hz, 1H), 4.09-4.11 (m, 1H), 4.33 (d,J=9.16 Hz, 1H), 4.43 (d, J=11.29 Hz, 1H), 4.54-4.57 (m, 1H), 5.12 (d,J=10.07 Hz, 1H), 5.30 (d, J=17.40 Hz, 1H), 5.73-5.80 (m, 1H), 5.88 (s,1H), 7.33 (d, J=5.49 Hz, 1H), 7.53 (m, 1H), 7.71 (t, J=6.87 Hz, 1H),7.81 (d, J=7.93 Hz, 1H), 7.97 (d, J=5.80 Hz, 1H), 8.19 (d, J=7.63 Hz,1H); MS m/z 698 (MH⁺).

Example 126 Preparation of Compound 126

Compound 126 was prepared by following Scheme 1 of Example 120 exceptthat 2-fluoroethyl chloroformate was used in place of p-tolylchloroformate in step 2.

Step 2:

Modifications: 24 mg (0.19 mmol) 2-fluoroethyl chloroformate used, 58.9mg product obtained as a yellow glassy solid (59.8% yield): ¹H NMR(CD₃OD) δ 1.05 (d, J=2.14 Hz, 9H), 1.07-1.09 (m, 2H), 1.22-1.27 (m, 2H),1.42-1.45 (m, 1H), 1.87-1.90 (m, 1H), 2.24 (q, J=8.75 Hz, 1H), 2.28-2.33(m, 1H), 2.63 (dd, J=13.43, 6.41 Hz, 1H), 2.92-2.96 (m, 1H), 3.92-4.10(m, 3H), 4.31-4.37 (m, 2H), 4.42-4.46 (m, 2H), 4.54-4.57 (m, 1H), 5.12(d, J=10.38 Hz, 1H), 5.29 (d, J=17.09 Hz, 1H), 5.71-5.79 (m, 1H), 5.88(s, 1H), 7.33 (d, J=5.80 Hz, 1H), 7.55 (t, J=7.17 Hz, 1H), 7.71 (m, 1H),7.81 (d, J=7.93 Hz, 1H), 7.96 (d, J=5.80 Hz, 1H), 8.19 (d, J=7.63 Hz,1H); MS m/z 674 (MH⁺).

Example 127 Preparation of Compound 127

Compound 127 was prepared by following Scheme 1 of Example 120 exceptthat 2-methoxyphenyl chloroformate was used in place of p-tolylchloroformate in step 2.

Step 2:

Modifications: 35 mg (0.19 mmol) 2-methoxyphenyl chloroformate used,97.6 mg product obtained as a sticky yellow oil (91.0% yield): MS m/z734 (MH⁺).

Example 128 Preparation of Compound 128

Compound 128 was prepared by following Scheme 1 of Example 120 exceptthat 2-(−)-(1R)-menthyl chloroformate was used in place of p-tolylchloroformate in step 2.

Step 2:

Modifications: 42 mg (0.19 mmol) (−)-(1R)-menthyl chloroformate used,69.1 mg product obtained as a white glassy solid (61.7% yield): MS m/z766 (MH⁺).

Example 129 Preparation of Compound 129

Compound 129 was prepared by following Scheme 1 of Example 120 exceptthat hexyl chloroformate was used in place of p-tolyl chloroformate instep 2.

Step 2:

Modifications: 31 mg (0.19 mmol) hexyl chloroformate used, 66.7 mgproduct obtained as a yellow glassy solid (64.1% yield): ¹H NMR (CD₃OD)δ 0.87-0.99 (m, H), 1.05 (s, 9H), 1.07-1.09 (m, 2H), 1.22-1.28 (m, 6H),1.43-1.48 (m, 3H), 1.88 (dd, J=8.24, 5.49 Hz, 1H), 2.24 (q, J=8.85 Hz,1H), 2.28-2.33 (m, 1H), 2.63 (dd, J=14.34, 7.63 Hz, 1H), 2.92-2.97 (m,1H), 3.72 (dt, J=10.61, 6.60 Hz, 1H), 3.81-3.86 (m, 1H), 4.10 (dd,J=11.60, 3.36 Hz, 1H), 4.32 (d, J=8.85 Hz, 1H), 4.43 (d, J=11.90 Hz,1H), 4.55 (dd, J=9.77, 7.32 Hz, 1H), 5.13 (d, J=10.38 Hz, 1H), 5.30 (d,J=17.09 Hz, 1H), 5.76 (ddd, J=17.09, 10.07, 9.16 Hz, 1H), 5.89 (s, 1H),7.33 (d, J=5.80 Hz, 1H), 7.54 (t, J=7.48 Hz, 1H), 7.69-7.72 (m, 1H),7.81 (d, J=8.24 Hz, 1H), 7.97 (d, J=6.10 Hz, 1H), 8.20 (d, J=8.24 Hz,1H); MS m/z 712 (MH⁺).

Example 130 Preparation of Compound 130

Step 1:

A solution of Compound 23 (see Example 23) (1.50 g, 2.19 mmol) in DCM(50 mL) and trifluoroacetic acid (50 mL) was stirred for 3 h at rt. Themixture was concentrated in vacuo to a viscous residue, and was thendissolved in 1,2-dichloroethane and again concentrated in vacuo to givethe desired bis-trifluoroacetic acid salt product as an off-white glassysolid (quantitative). The material was used directly in the next stepwithout purification.

Step 2:

A mixture of the product from step 1 (118 mg, 0.146 mmol), tert-butylacetic acid (22 mg, 0.19 mmol), HATU (72 mg, 0.19 mmol) andN-methylmorpholine (59 mg, 0.58 mmol) in 1,2-dichloroethane was stirredfor 24 h at rt. The reaction mixture was washed with pH=4 buffersolution (3×3 mL), and the washes were back-extracted with1,2-dichloroethane (3 mL). The organic phases were combined andconcentrated in vacuo. The crude product was then dissolved in MeOH andpurified by reverse phase preparative HPLC to give the title compound(Compound 130) as a slightly yellow glassy solid (43.4 mg, 43.5% yield):¹H NMR (CD₃OD) δ 0.82 (d, J=1.83 Hz, 9H), 1.06 (d, J=2.14 Hz, 9H),1.07-1.10 (m, 2H), 1.22-1.28 (m, 2H), 1.43-1.46 (m, 1H), 1.87-1.90 (m,1H), 1.99 (d, J=1.83 Hz, 2H), 2.20-2.26 (m, 1H), 2.27-2.33 (m, 1H),2.59-2.64 (m, 1H), 2.93-2.97 (m, 1H), 4.12-4.14 (m, 1H), 4.42 (d,J=11.60 Hz, 1H), 4.51-4.55 (m, 1H), 4.67 (dd, J=9.31, 1.98 Hz, 1H),5.11-5.14 (m, 1H), 5.29 (d, J=17.40 Hz, 1H), 5.72-5.80 (m, 1H), 5.89 (d,J=1.83 Hz, 1H), 7.32 (dd, J=5.80, 2.14 Hz, 1H), 7.52-7.55 (m, 1H),7.69-7.72 (m, 1H), 7.81 (d, J=8.24 Hz, 1H), 7.96 (dd, J=5.80, 1.83 Hz,1H), 8.17 (d, J=8.24 Hz, 1H); MS m/z 682 (MH⁺).

Example 131 Preparation of Compound 131

Compound 131 was prepared by following Scheme 1 of Example 130 exceptthat methoxyacetic acid was used in place of tert-butyl acetic acid instep 2.

Step 2:

Modifications: 17 mg (0.19 mmol) methoxyacetic acid used, 49.9 mgproduct obtained as a slightly yellow glassy solid (52.0% yield): ¹H NMR(CD₃OD) δ 1.05-1.08 (m, 11H), 1.24-1.26 (m, 2H), 1.45 (ddd, J=9.31,5.34, 3.66 Hz, 1H), 1.88 (ddd, J=8.39, 5.19, 3.81 Hz, 1H), 2.21-2.27 (m,1H), 2.29-2.35 (m, 1H), 2.60-2.64 (m, 1H), 2.91-2.97 (m, 1H), 3.34 (d,J=3.66 Hz, 3H), 3.69 (dd, J=15.26, 3.66 Hz, 1H), 3.81-3.85 (m, 1H), 4.15(dt, J=11.67, 3.62 Hz, 1H), 4.35 (d, J=11.90 Hz, 1H), 4.55 (ddd,J=10.30, 6.94, 3.20 Hz, 1H), 4.66 (dd, J=9.61, 3.51 Hz, 1H), 5.11-5.14(m, 1H), 5.28-5.32 (m, 1H), 5.73-5.81 (m, 1H), 5.90 (d, J=3.36 Hz, 1H),7.33 (dd, J=5.65, 3.20 Hz, 1H), 7.54-7.58 (m, 1H), 7.69-7.73 (m, 1H),7.80-7.82 (m, 1H), 7.95-7.97 (m, 1H), 8.15 (dd, J=8.39, 2.59 Hz, 1H); MSm/z 656 (MH⁺).

Example 132 Preparation of Compound 132

Compound 132 was prepared by following Scheme 1 of Example 130 exceptthat methoxypropionic acid was used in place of tert-butyl acetic acidin step 2.

Step 2:

Modifications: 20 mg (0.19 mmol) methoxypropionic acid used, 50.0 mgproduct obtained as a yellow glassy solid (51.1% yield): ¹H NMR (CD₃OD)δ 1.06 (d, J=1.83 Hz, 9H), 1.07-1.09 (m, 2H), 1.23-1.27 (m, 2H), 1.44(ddd, J=9.38, 5.26, 1.83 Hz, 1H), 1.87-1.90 (m, 1H), 2.21-2.27 (m, 1H),2.29-2.33 (m, 2H), 2.40-2.46 (m, 1H), 2.59-2.64 (m, 1H), 2.92-2.97 (m,1H), 3.25 (d, J=1.83 Hz, 3H), 3.45-3.54 (m, 2H), 4.12-4.16 (m, 1H), 4.37(d, J=11.60 Hz, 1H), 4.52-4.55 (m, 1H), 4.65 (dd, J=9.16, 1.83 Hz, 1H),5.12 (d, J=10.38 Hz, 1H), 5.30 (d, J=17.40 Hz, 1H), 5.72-5.80 (m, 1H),5.89 (s, 1H), 7.33 (dd, J=5.65, 1.98 Hz, 1H), 7.54-7.58 (m, 1H),7.69-7.73 (m, 1H), 7.81 (d, J=8.24 Hz, 1H), 7.96 (dd, J=6.10, 1.83 Hz,1H), 8.18 (d, J=8.24 Hz, 1H); MS m/z 670 (MH⁺).

Example 133 Preparation of Compound 133

Compound 133 was prepared by following Scheme 1 of Example 130 exceptthat (S)-1,4-benzodioxane-2-carboxylic acid was used in place oftert-butyl acetic acid in step 2.

Step 2:

Modifications: 35 mg (0.19 mmol) (S)-1,4-benzodioxane-2-carboxylic acidused, 54.0 mg product obtained as a slightly yellow glassy solid (49.5%yield): ¹H NMR (CD₃OD) δ 0.83 (d, J=3.36 Hz, 9H), 1.05-1.08 (m, 2H),1.20-1.25 (m, 1H), 1.26-1.31 (m, 1H), 1.45-1.49 (m, 1H), 1.86-1.90 (m,1H), 2.21-2.25 (m, 1H), 2.28-2.34 (m, 1H), 2.59-2.65 (m, 1H), 2.90-2.94(m, 1H), 4.12-4.17 (m, 2H), 4.32 (d, J=11.90 Hz, 1H), 4.35-4.39 (m, 1H),4.55-4.61 (m, 3H), 5.11-5.14 (m, 1H), 5.28-5.32 (m, 1H), 5.75-5.83 (m,1H), 5.90 (d, J=3.66 Hz, 1H), 6.80-6.89 (m, 3H), 7.03-7.07 (m, 1H),7.32-7.34 (m, 1H), 7.55-7.58 (m, 1H), 7.68-7.72 (m, 1H), 7.80-7.82 (m,1H), 7.96-7.98 (m, 1H), 8.15-8.18 (m, 1H); MS m/z 746 (MH⁺).

Example 134 Preparation of Compound 134

Step 1:

A mixture of the product from Example 11, Step 5 (100 mg, 0.172 mmol),N-α-tert-butoxycarbonyl-L-phenylglycine (45.3 mg, 0.180 mmol), HATU(84.9 mg, 0.223 mmol), and N-methylmorpholine (87.0 mg, 0.859 mmol) inDMF (1.0 mL) was stirred at rt for 18 h. The mixture was purifieddirectly by reverse phase preparative HPLC to give 29.7 mg (23.6% yield)of Compound 134 as a white powder: ¹H NMR (CD₃OD) δ 0.97-1.07 (m, 2H),1.12-1.17 (m, 1H), 1.22-1.32 (m, 2H), 1.38 (s, 9H), 1.90 (dd, J=8.09,5.34 Hz, 1H), 2.20-2.28 (m, 2H), 2.54 (dd, J=13.58, 6.56 Hz, 1H),2.85-2.89 (m, J=8.24 Hz, 1H), 3.50 (d, J=10.99 Hz, 1H), 3.93 (s, 3H),4.11 (d, J=11.60 Hz, 1H), 4.63 (dd, J=9.46, 7.32 Hz, 1H), 5.13 (dd,J=10.38, 1.53 Hz, 1H), 5.32 (d, J=17.09 Hz, 1H), 5.47 (s, 1H), 5.74-5.84(m, 2H), 7.16-7.19 (m, 1H), 7.25 (d, J=5.80 Hz, 1H), 7.32-7.43 (m, 6H),7.86 (d, J=5.80 Hz, 1H), 8.13 (d, J=9.16 Hz, 1H); MS m/z 734 (MH⁺).

Example 135 Preparation of Compound 135

Compound 135 was prepared by following Scheme 1 of Example 134 exceptthat N-α-tert-butoxycarbonyl-erythro-DL-β-methylphenylalanine was usedin place of N-α-tert-butoxycarbonyl-L-phenylglycine in step 1. Compound135 was prepared from a mixture of N-α-tert-butoxycarbonyl-erythro DLβ-methylphenylalanine and the resulting two diastereomers were separatedby reverse phase preparative HPLC. This compound is the single isomerwhich eluted first from the preparative HPLC column. The exactstereochemistry at the β-methyl phenylalanine portion of the molecule isunknown.

Step 1:

Modifications: 50.4 mg (0.180 mmol)N-α-tert-butoxycarbonyl-erythro-DL-β-methylphenylalanine used, 29.7 mgproduct obtained as a white powder (22.7% yield): ¹H NMR (CD₃OD) δ 1.11(d, J=7.93 Hz, 2H), 1.15 (d, J=6.10 Hz, 3H), 1.24-1.32 (m, 11H), 1.44(dd, J=9.16, 5.19 Hz, 1H), 1.90-1.94 (m, 1H), 2.25-2.29 (m, 1H), 2.36(t, J=13.28 Hz, 1H), 2.62 (dd, J=13.58, 7.17 Hz, 1H), 2.98-3.02 (m, 1H),3.20-3.24 (m, 1H), 3.91 (s, 3H), 4.11 (dd, J=11.60, 3.05 Hz, 1H), 4.51(d, J=10.68 Hz, 1H), 4.57 (dd, J=10.07, 7.32 Hz, 1H), 4.63 (d, J=12.21Hz, 1H), 5.14 (d, J=10.07 Hz, 1H), 5.32 (d, J=16.79 Hz, 1H), 5.76-5.84(m, 1H), 5.88 (s, 1H), 7.08 (dd, J=8.70, 1.68 Hz, 1H), 7.16-7.18 (m,2H), 7.23-7.27 (m, 5H), 7.89 (d, J=5.80 Hz, 1H), 8.09 (d, J=9.46 Hz,1H); MS m/z 762 (MH⁺).

Example 136 Preparation of Compound 136

Compound 136 was prepared by following Scheme 1 of Example 134 exceptthat N-α-tert-butoxycarbonyl-erythro-DL-β-methylphenylalanine was usedin place of N-α-tert-butoxycarbonyl-L-phenylglycine in step 1. Compound136 was prepared from a mixture of N-tert-butoxycarbonyl-erythro DLβ-methylphenylalanine and the resulting two diastereomers were separatedby reverse phase preparative HPLC. This compound is the single isomerwhich eluted second from the preparative HPLC column. The exactstereochemistry at the β-methyl phenylalanine portion of the molecule isunknown.

Step 1:

Modifications: 50.4 mg (0.180 mmol)N-α-tert-butoxycarbonyl-erythro-DL-β-methylphenylalanine used, 26.3 mgproduct obtained as a white powder (20.1% yield): ¹H NMR (CD₃OD) δ 1.04(s, 1H), 1.13 (d, J=6.71 Hz, 3H), 1.12-1.17 (m, 2H), 1.30 (s, 9H),1.33-1.36 (m, 1H), 1.41 (dd, J=9.46, 5.19 Hz, 1H), 1.87 (dd, J=7.78,5.34 Hz, 1H), 2.29 (q, J=8.85 Hz, 1H), 2.36 (ddd, J=13.81, 9.99, 4.27Hz, 1H), 2.54 (dd, J=13.58, 7.17 Hz, 1H), 3.00-3.04 (m, 1H), 3.05-3.08(m, 1H), 3.80 (d, J=11.90 Hz, 1H), 3.94 (s, 3H), 4.10 (dd, J=12.05, 3.81Hz, 1H), 4.53-4.57 (m, 1H), 4.59 (d, J=8.24 Hz, 1H), 5.14 (d, J=10.38Hz, 1H), 5.34 (d, J=17.09 Hz, 1H), 5.78-5.85 (m, 2H), 6.75 (t, J=7.32Hz, 1H), 7.03 (t, J=7.48 Hz, 2H), 7.12 (s, 1H), 7.14 (s, 1H), 7.19 (dd,J=9.31, 1.68 Hz, 1H), 7.22 (s, 1H), 7.28 (d, J=6.10 Hz, 1H), 7.89 (d,J=5.80 Hz, 1H), 8.03 (d, J=8.85 Hz, 1H); MS m/z 762 (MH⁺).

Example 137 Preparation of Compound 137

Step 1:

A mixture of the product from Example 11, Step 5 (100 mg, 0.172 mmol),N-α-tert-butoxycarbonyl-L-aspartic acid 4-benzyl ester (59.5 mg, 0.180mmol), HATU (84.9 mg, 0.223 mmol), and N-methylmorpholine (87.0 mg,0.859 mmol) in DCM (3.0 mL) was stirred at rt for 18 h. The reactionmixture was washed with pH=4 buffer solution (3×3 mL), and the washeswere back-extracted with DCM (3 mL). The organic phases were combinedand concentrated in vacuo. The crude product was then dissolved in MeOHand purified by reverse phase preparative HPLC to give Compound 137 as aslightly off-white glassy solid (26.0 mg, 18.8% yield): ¹H NMR (CD₃OD)80.95-1.01 (m, 2H), 1.16 (s, 9H), 1.22-1.29 (m, 2H), 1.44 (dd, J=9.46,5.19 Hz, 1H), 1.86 (dd, J=7.93, 5.19 Hz, 1H), 2.26 (q, J=8.85 Hz, 1H),2.32-2.37 (m, 1H), 2.61 (dd, J=13.73, 7.32 Hz, 1H), 2.66 (dd, J=16.48,6.10 Hz, 1H), 2.89 (ddd, J=12.67, 8.09, 4.88 Hz, 1H), 3.05 (dd, J=16.63,8.39 Hz, 1H), 3.92 (s, 3H), 4.04-4.07 (m, 1H), 4.47 (d, J=11.90 Hz, 1H),4.52-4.56 (m, 1H), 4.75 (dd, J=8.24, 6.41 Hz, 1H), 5.12-5.14 (m, 1H),5.14 (s, 2H), 5.31 (d, J=17.09 Hz, 1H), 5.75-5.82 (m, 2H), 7.12 (d,J=9.16 Hz, 1H), 7.18 (d, J=2.14 Hz, 1H), 7.24 (d, J=5.80 Hz, 1H),7.31-7.33 (m, 1H), 7.36 (t, J=7.32 Hz, 2H), 7.39 (s, 1H), 7.41 (s, 1H),7.88 (d, J=6.10 Hz, 1H), 8.10 (d, J=9.16 Hz, 1H); MS m/z 806 (MH⁺).

Example 138 Preparation of Compound 138

Compound 138 was prepared by following Scheme 1 of Example 137 exceptthat N-tert-butoxycarbonyl-L-aspartic acid 4-methyl ester was used inplace of N-α-tert-butoxycarbonyl-L-aspartic acid 4-benzyl ester in step1.

Step 1:

Modifications: 45.5 mg (0.180 mmol) N-tert-butoxycarbonyl-L-asparticacid 4-methyl ester used, 93.5 mg product obtained as an off-whiteglassy solid (74.6% yield): ¹H NMR (CD₃OD) δ 1.07-1.09 (m, 2H), 1.17 (s,9H), 1.20-1.29 (m, 2H), 1.41-1.44 (m, 1H), 1.84-1.86 (m, 1H), 2.26 (q,J=8.85 Hz, 1H), 2.33-2.38 (m, 1H), 2.58-2.64 (m, 2H), 2.92-3.02 (m, 2H),3.69 (s, 3H), 3.92 (s, 3H), 4.15 (dd, J=11.44, 2.29 Hz, 1H), 4.49-4.56(m, 2H), 4.72-4.76 (m, 1H), 5.13 (d, J=10.38 Hz, 1H), 5.32 (d, J=17.09Hz, 1H), 5.74-5.82 (m, 1H), 5.87 (s, 1H), 7.12 (d, J=9.16 Hz, 1H), 7.18(s, 1H), 7.24 (d, J=5.80 Hz, 1H), 7.88 (dd, J=5.80, 0.92 Hz, 1H), 8.10(d, J=8.85 Hz, 1H); MS m/z 730 (MH⁺).

Example 139 Preparation of Compound 139

Compound 139 was prepared by following Scheme 1 of Example 137 exceptthat N-tert-butoxycarbonyl-L-aspartic acid 4-tert-butyl ester was usedin place of N-α-tert-butoxycarbonyl-L-aspartic acid 4-benzyl ester instep 1.

Step 1:

Modifications: 52.2 mg (0.180 mmol) N-tert-butoxycarbonyl-L-asparticacid 4-tert-butyl ester used, 125 mg product obtained as an off-whiteglassy solid (99.8% yield):

¹H NMR (CD₃OD) δ 1.08-1.10 (m, 2H), 1.17 (s, 9H), 1.21-1.29 (m, 2H),1.46 (s, H), 1.82 (dd, J=7.78, 5.34 Hz, 1H), 2.26 (q, J=8.75 Hz, 1H),2.32-2.38 (m, 1H), 2.51 (dd, J=16.33, 7.17 Hz, 1H), 2.63 (dd, J=14.04,7.02 Hz, 1H), 2.89 (dd, J=16.48, 7.63 Hz, 1H), 2.92-2.98 (m, 1H), 3.92(s, 3H), 4.15 (dd, J=11.60, 3.05 Hz, 1H), 4.50-4.57 (m, 2H), 4.70-4.75(m, 1H), 5.13 (dd, J=10.38, 1.53 Hz, 1H), 5.32 (d, J=17.40 Hz, 1H),5.76-5.83 (m, 1H), 5.87 (s, 1H), 7.13 (dd, J=8.85, 1.53 Hz, 1H), 7.18(d, J=2.14 Hz, 1H), 7.25 (d, J=5.80 Hz, 1H), 7.88 (d, J=5.80 Hz, 1H),8.10 (d, J=9.16 Hz, 1H); MS m/z 772 (MH⁺).

Example 140 Preparation of Compound 140

Compound 138 (50.0 mg, 0.0685 mmol) was dissolved in a mixture of THF (1mL), MeOH (1 mL), and 1.0M aqueous NaOH (0.137 mL, 0.137 mmol). After 3h, the reaction mixture was neutralized by the addition of 1.0M aqueousHCl (0.137 mL, 0.137 mmol). The crude mixture was concentrated in vacuo,then pH=4 buffer solution (3 mL) and DCM (3 mL) were added and themixture was shaken. The layers were separated and the aqueous layer wasfurther extracted with DCM (2×1 mL). The organic phases were combined,dried over anhydrous MgSO₄, filtered and concentrated in vacuo to give48.0 mg (97.9% yield) of Compound 140 as an off white glassy solid: ¹HNMR (CD₃OD) δ 1.04-1.07 (m, 2H), 1.17 (s, 9H), 1.22-1.25 (m, 2H), 1.38(dd, J=9.31, 5.34 Hz, 1H), 1.78 (dd, J=7.78, 5.34 Hz, 1H), 2.30 (q,J=8.75 Hz, 1H), 2.37-2.42 (m, 1H), 2.48 (dd, J=15.87, 4.58 Hz, 1H), 2.72(dd, J=13.12, 7.63 Hz, 1H), 2.88 (dd, J=15.72, 10.53 Hz, 1H), 2.90-2.95(m, 1H), 3.92 (s, 3H), 4.21 (dd, J=11.44, 2.90 Hz, 1H), 4.57 (t, J=8.70Hz, 1H), 4.62-4.65 (m, 2H), 5.10 (d, J=10.68 Hz, 1H), 5.34 (d, J=17.09Hz, 1H), 5.69-5.76 (m, 1H), 5.83 (s, 1H), 7.10 (d, J=9.16 Hz, 1H), 7.17(s, 1H), 7.23 (d, J=6.10 Hz, 1H), 7.88 (d, J=6.10 Hz, 1H), 8.12 (d,J=8.85 Hz, 1H); MS m/z 716 (MH⁺).

Example 141 Preparation of Compound 141

Step 1:

The product of Example 55, Step 1 (65 mg, 0.0947 mmol),N,N′-disuccinimidyl carbonate (41.0 mg, 0.142 mmol) andN,N-diisopropylethylamine (30.6 mg, 0.237 mmol) were combined withanhydrous THF (1 mL) and the resulting suspension was heated to 80° C.in a microwave reactor for 15 min. Upon cooling to rt, this crudemixture was used directly in the next step.

Step 2:

The crude reaction mixture from step 1 was treated with a mixture ofL-valine methyl ester hydrochloride (159 mg, 0.947 mmol) andN,N-diisopropylethylamine (122 mg, 0.947 mmol) in anhydrous THF (2 mL).The resulting mixture was stirred for 18 h at rt. Solvent was removed invacuo, and the residue was taken up in DCM (2 mL) and washed with pH=4buffer solution (3×2 mL). The buffer washes were combined andback-extracted with DCM (2 mL). The combined DCM phases wereconcentrated in vacuo, and the resulting residue was dissolved in MeOHand purified by reverse phase preparative HPLC to give 38.1 mg (52.2%yield) of Compound 141 as a white powder: ¹H NMR (CD₃OD) δ 0.83 (dd,J=6.87, 3.81 Hz, 6H), 1.06 (s, 11H), 1.21-1.26 (m, 2H), 1.41 (dd,J=9.46, 5.49 Hz, 1H), 1.87 (dd, J=8.09, 5.34 Hz, 1H), 1.95-2.02 (m, 1H),2.21 (q, J=8.85 Hz, 1H), 2.29 (ddd, J=13.89, 9.92, 4.27 Hz, 1H), 2.60(dd, J=13.73, 7.02 Hz, 1H), 2.91-2.97 (m, 1H), 3.67 (s, 3H), 3.93 (s,3H), 4.00 (d, J=5.49 Hz, 1H), 4.09 (dd, J=11.90, 3.97 Hz, 1H), 4.40-4.43(m, 2H), 4.52 (dd, J=10.07, 7.02 Hz, 1H), 5.11 (dd, J=10.38, 1.53 Hz,1H), 5.28 (dd, J=17.09, 1.22 Hz, 1H), 5.75 (ddd, J=17.17, 10.15, 9.00Hz, 1H), 5.83 (s, 1H), 7.11 (dd, J=9.16, 2.44 Hz, 1H), 7.18 (d, J=2.44Hz, 1H), 7.24 (d, J=5.80 Hz, 1H), 7.87 (d, J=6.10 Hz, 1H), 8.10 (d,J=8.85 Hz, 1H); MS m/z 771 (MH⁺).

Example 142 Preparation of Compound 142

Compound 142 was prepared by following Scheme 1 of Example 141 exceptthat D-valine methyl ester hydrochloride was used in place of L-valinemethyl ester hydrochloride in step 2.

Step 2:

Modifications: 159 mg (0.947 mmol) D-valine methyl ester hydrochlorideused, 23.0 mg product obtained as a white powder (31.5% yield): ¹H NMR(CD₃OD) δ 0.88 (dd, J=13.89, 6.87 Hz, 6H), 1.06 (s, 9H), 1.07-1.09 (m,2H), 1.23-1.27 (m, 2H), 1.41 (dd, J=9.46, 5.49 Hz, 1H), 1.88 (dd,J=7.93, 5.49 Hz, 1H), 2.01-2.07 (m, 1H), 2.23 (q, J=9.05 Hz, 1H), 2.31(ddd, J=14.11, 9.99, 4.27 Hz, 1H), 2.63 (dd, J=13.89, 7.17 Hz, 1H),2.93-2.98 (m, 1H), 3.62 (s, 3H), 3.96 (s, 3H), 4.03 (d, J=5.19 Hz, 1H),4.09 (dd, J=11.75, 3.81 Hz, 1H), 4.38 (s, 1H), 4.48-4.54 (m, 2H), 5.12(dd, J=10.38, 1.22 Hz, 1H), 5.29 (dd, J=17.24, 1.07 Hz, 1H), 5.70-5.77(m, 1H), 5.83 (s, 1H), 7.22 (dd, J=9.00, 2.59 Hz, 1H), 7.25 (d, J=2.44Hz, 1H), 7.35 (d, J=6.10 Hz, 1H), 7.87 (d, J=6.10 Hz, 1H), 8.16 (d,J=9.16 Hz, 1H); MS m/z 771 (MH⁺).

Example 143 Preparation of Compound 143

Compound 143 was prepared by following Scheme 1 of Example 137 exceptthat N-tert-butoxycarbonyl-L-cyclohexylglycine was used in place ofN-α-tert-butoxycarbonyl-L-aspartic acid 4-benzyl ester in step 1.

Step 1:

Modifications: 46.2 mg (0.180 mmol)N-tert-butoxycarbonyl-L-cyclohexylglycine used, 93.9 mg product obtainedas a white powder (73.8% yield): ¹H NMR (CD₃OD) δ 1.04-1.08 (dd, J=7.78,2.29 Hz, 4H), 1.19-1.26 (m, 4H), 1.25 (s, 9H), 1.41 (dd, J=9.46, 5.19Hz, 1H), 1.63-1.82 (m, 7H), 1.88 (dd, J=7.93, 5.49 Hz, 1H), 2.22 (q,J=9.05 Hz, 1H), 2.32-2.37 (m, 1H), 2.59 (dd, J=13.58, 6.87 Hz, 1H),2.91-2.96 (m, 1H), 3.92 (s, 3H), 4.05 (dd, J=11.75, 3.20 Hz, 1H), 4.09(d, J=8.85 Hz, 1H), 4.47 (d, J=11.90 Hz, 1H), 4.53 (dd, J=10.22, 7.17Hz, 1H), 5.11 (d, J=10.38 Hz, 1H), 5.29 (d, J=16.79 Hz, 1H), 5.79 (ddd,J=16.86, 9.92, 9.54 Hz, 1H), 5.84 (s, 1H), 7.10 (d, J=8.85 Hz, 1H), 7.17(d, J=1.53 Hz, 1H), 7.24 (d, J=5.80 Hz, 1H), 7.88 (d, J=6.10 Hz, 1H),8.09 (d, J=8.85 Hz, 1H); MS m/z 740 (MH⁺).

Example 144 Preparation of Compound 144

Compound 144 was prepared by following Scheme 1 of Example 141 exceptthat the scale was increased and that glycine methyl ester hydrochloridewas used in place of L-valine methyl ester hydrochloride in step 2.

Step 1:

Modifications: 100 mg (0.146 mmol) of the product of Example 55, Step 1;62.2 mg (0.219 mmol) N,N′-disuccinimidyl carbonate, 47.0 mg (0.364 mmol)N,N-diisopropylethylamine used.

Step 2:

Modifications: 183 mg (1.46 mmol) glycine methyl ester hydrochloride,188 mg (1.46 mmol) N,N-diisopropylethylamine used, 56.3 mg productobtained as a white powder (52.9% yield): ¹H NMR (CD₃OD) δ 1.01-1.04 (m,2H), 1.05 (s, 9H), 1.17-1.21 (m, 2H), 1.40 (dd, J=9.46, 5.49 Hz, 1H),1.85 (dd, J=7.78, 5.34 Hz, 1H), 2.18 (q, J=8.55 Hz, 1H), 2.33 (ddd,J=13.89, 9.92, 4.27 Hz, 1H), 2.61 (dd, J=13.73, 7.32 Hz, 1H), 2.89-2.94(m, 1H), 3.65 (s, 3H), 3.69-3.77 (m, 2H), 3.92 (s, 3H), 4.10 (dd,J=11.75, 3.81 Hz, 1H), 4.40-4.42 (m, 2H), 4.53 (dd, J=9.92, 7.17 Hz,1H), 5.08 (d, J=10.38 Hz, 1H), 5.26 (d, J=17.09 Hz, 1H), 5.77 (ddd,J=17.09, 10.22, 9.00 Hz, 1H), 5.83 (s, 1H), 7.13 (dd, J=8.85, 2.44 Hz,1H), 7.17 (s, 1H), 7.23 (d, J=5.80 Hz, 1H), 7.87 (d, J=5.80 Hz, 1H),8.09 (d, J=8.85 Hz, 1H); MS m/z 729 (MH⁺).

Example 145 Preparation of Compound 145

Compound 145 was prepared by following Scheme 1 of Example 141 exceptthat the scale was increased and that L-alanine methyl esterhydrochloride was used in place of L-valine methyl ester hydrochloridein step 2.

Step 1:

Modifications: 100 mg (0.146 mmol) of the product of Example 55, Step 1;62.2 mg (0.219 mmol) N,N′-disuccinimidyl carbonate, 47.0 mg (0.364 mmol)N,N-diisopropylethylamine used.

Step 2:

Modifications: 203 mg (1.46 mmol) L-alanine methyl ester hydrochloride,188 mg (1.46 mmol) N,N-diisopropylethylamine used, 64.3 mg productobtained as a white powder (59.3% yield): ¹H NMR (CD₃OD) δ 0.97-1.02 (m,2H), 1.05 (s, 9H), 1.19 (d, J=7.02 Hz, 3H), 1.18-1.22 (m, 2H), 1.41 (dd,J=9.46, 5.19 Hz, 1H), 1.86 (dd, J=8.09, 5.34 Hz, 1H), 2.19 (q, J=8.85Hz, 1H), 2.32 (ddd, J=13.81, 9.84, 4.43 Hz, 1H), 2.61 (dd, J=13.73, 7.02Hz, 1H), 2.93 (ddd, J=12.82, 8.09, 4.73 Hz, 1H), 3.65 (s, 3H), 3.93 (s,3H), 3.99 (q, J=7.22 Hz, 1H), 4.08 (dd, J=11.75, 3.81 Hz, 1H), 4.38 (s,1H), 4.42 (d, J=11.60 Hz, 1H), 4.53 (dd, J=10.07, 7.32 Hz, 1H), 5.09(dd, J=10.38, 1.53 Hz, 1H), 5.27 (dd, J=17.09, 1.22 Hz, 1H), 5.77 (ddd,J=17.09, 10.07, 9.16 Hz, 1H), 5.82 (s, 1H), 7.13 (dd, J=9.16, 2.44 Hz,1H), 7.18 (d, J=2.44 Hz, 1H), 7.24 (d, J=5.80 Hz, 1H), 7.87 (d, J=5.80Hz, 1H), 8.10 (d, J=9.16 Hz, 1H); MS m/z 743 (MH⁺).

Example 146 Preparation of Compound 146

Compound 147 was prepared by following Scheme 1 of Example 141 exceptthat the scale was increased and that L-tert-leucine methyl esterhydrochloride was used in place of L-valine methyl ester hydrochloridein step 2.

Step 1:

Modifications: 100 mg (0.146 mmol) of the product of Example 55, Step 1;62.2 mg (0.219 mmol) N,N′-disuccinimidyl carbonate, 47.0 mg (0.364 mmol)N,N-diisopropylethylamine used.

Step 2:

Modifications: 265 mg (1.46 mmol) L-tert-leucine methyl esterhydrochloride, 188 mg (1.46 mmol) N,N-diisopropylethylamine used, 68.3mg product obtained as a white powder (59.6% yield): ¹H NMR (CD₃OD) δ0.88 (s, 9H), 0.97-1.03 (m, 2H), 1.06 (s, 9H), 1.18-1.24 (m, 2H), 1.41(dd, J=9.46, 5.49 Hz, 1H), 1.86 (dd, J=7.93, 5.49 Hz, 1H), 2.19 (q,J=8.75 Hz, 1H), 2.30 (ddd, J=13.89, 10.07, 4.43 Hz, 1H), 2.60 (dd,J=13.73, 7.32 Hz, 1H), 2.91-2.96 (m, 1H), 3.65 (s, 3H), 3.91 (s, 1H),3.93 (s, 3H), 4.09 (dd, J=11.60, 3.97 Hz, 1H), 4.38 (s, 1H), 4.43 (d,J=11.60 Hz, 1H), 4.51 (dd, J=10.07, 7.32 Hz, 1H), 5.10 (dd, J=10.38,1.53 Hz, 1H), 5.27 (dd, J=17.24, 1.37 Hz, 1H), 5.76 (ddd, J=17.09,10.07, 9.16 Hz, 1H), 5.82 (s, 1H), 7.12 (dd, J=9.16, 2.44 Hz, 1H), 7.18(d, J=2.14 Hz, 1H), 7.24 (d, J=6.10 Hz, 1H), 7.87 (d, J=5.80 Hz, 1H),8.11 (d, J=9.16 Hz, 1H); MS m/z 785 (MH⁺).

Example 147 Preparation of Compound 147

Compound 147 was prepared by following Scheme 1 of Example 141 exceptthat the scale was increased and that L-histidine methyl esterhydrochloride was used in place of L-valine methyl ester hydrochloridein step 2, and the amount of N,N-diisopropylethylamine used in step 2was doubled.

Step 1:

Modifications: 100 mg (0.146 mmol) of the product of Example 55, Step 1;62.2 mg (0.219 mmol) N,N′-disuccinimidyl carbonate, 47.0 mg (0.364 mmol)N,N-diisopropylethylamine used.

Step 2:

Modifications: 352 mg (1.46 mmol) L-histidine methyl esterhydrochloride, 377 mg (2.91 mmol) N,N-diisopropylethylamine used, 51.0mg product obtained as a white powder (43.2% yield): ¹H NMR (CD₃OD) δ1.04 (s, 11H), 1.20-1.22 (m, 2H), 1.41 (ddd, J=9.46, 5.34, 1.07 Hz, 1H),1.86-1.88 (m, 1H), 2.23 (q, J=8.75 Hz, 1H), 2.28-2.33 (m, 1H), 2.60 (dd,J=13.73, 7.02 Hz, 1H), 2.92 (d, J=6.41 Hz, 2H), 2.92-2.96 (m, 1H), 3.64(s, 3H), 3.91 (d, J=1.53 Hz, 3H), 4.04 (dd, J=11.90, 3.66 Hz, 1H), 4.35(s, 1H), 4.36-4.41 (m, 2H), 4.53 (dd, J=9.77, 7.63 Hz, 1H), 5.10 (d,J=10.38 Hz, 1H), 5.28 (d, J=17.40 Hz, 1H), 5.73-5.78 (m, 1H), 5.81 (s,1H), 6.82 (s, 1H), 7.06-7.09 (m, 1H), 7.15 (s, 1H), 7.23 (d, J=5.80 Hz,1H), 7.63 (s, 1H), 7.87 (dd, J=5.80, 1.22 Hz, 1H), 8.08 (d, J=9.16 Hz,1H); MS m/z 809 (MH⁺).

Example 148 Preparation of Compound 148

Compound 148 was prepared by following Scheme 1 of Example 141 exceptthat the scale was increased and that L-valine ethyl ester hydrochloridewas used in place of L-valine methyl ester hydrochloride in step 2.

Step 1:

Modifications: 100 mg (0.146 mmol) of the product of Example 55, Step 1;62.2 mg (0.219 mmol) N,N′-disuccinimidyl carbonate, 47.0 mg (0.364 mmol)N,N-diisopropylethylamine used.

Step 2:

Modifications: 265 mg (1.46 mmol) L-valine ethyl ester hydrochloride,188 mg (1.46 mmol) N,N-diisopropylethylamine used, 69.2 mg productobtained as a white powder (60.4% yield): ¹H NMR (CD₃OD) δ 0.83 (dd,J=6.71, 5.19 Hz, 6H), 1.01-1.03 (m, 2H), 1.06 (s, 9H), 1.17-1.22 (m,2H), 1.23 (t, J=7.17 Hz, 3H), 1.40 (dd, J=9.46, 5.19 Hz, 1H), 1.86 (dd,J=8.09, 5.34 Hz, 1H), 1.95-2.02 (m, 1H), 2.18 (q, J=9.05 Hz, 1H), 2.33(ddd, J=13.89, 9.92, 4.27 Hz, 1H), 2.60 (dd, J=13.89, 7.17 Hz, 1H), 2.92(ddd, J=12.82, 8.09, 4.73 Hz, 1H), 3.93 (s, 3H), 3.98 (d, J=5.19 Hz,1H), 4.08-4.17 (m, 3H), 4.41 (s, 1H), 4.41-4.43 (m, 1H), 4.52 (dd,J=10.07, 7.32 Hz, 1H), 5.09 (dd, J=10.38, 1.53 Hz, 1H), 5.26 (dd,J=17.09, 1.22 Hz, 1H), 5.77 (ddd, J=17.09, 10.07, 9.16 Hz, 1H), 5.82 (s,1H), 7.12 (dd, J=9.16, 2.44 Hz, 1H), 7.18 (d, J=2.44 Hz, 1H), 7.23 (d,J=6.10 Hz, 1H), 7.87 (d, J=6.10 Hz, 1H), 8.10 (d, J=9.16 Hz, 1H); MS m/z785 (MH⁺).

Example 149 Preparation of Compound 149

Compound 149 was prepared by following Scheme 1 of Example 137 exceptthat N-tert-butoxycarbonyl-L-cyclopentylglycine dicyclohexylamine saltwas used in place of N-α-tert-butoxycarbonyl-L-aspartic acid 4-benzylester in step 1.

Step 1:

Modifications: 76.2 mg (0.180 mmol)N-tert-butoxycarbonyl-L-cyclopentylglycine dicyclohexylamine salt used,111 mg product obtained as a white powder (89.3% yield): ¹H NMR (CD₃OD)δ 0.98 (d, J=8.24 Hz, 2H), 1.15-1.18 (m, 2H), 1.24 (s, 9H), 1.29-1.32(m, J=18.01 Hz, 2H), 1.38-1.40 (m, 1H), 1.44 (dd, J=4.88, 1.53 Hz, 1H),1.49-1.55 (m, 2H), 1.62-1.67 (m, 2H), 1.74-1.80 (m, 1H), 1.85-1.88 (m,1H), 2.16 (q, J=8.75 Hz, 1H), 2.21-2.26 (m, 1H), 2.42 (t, J=11.90 Hz,1H), 2.60-2.64 (m, 1H), 2.89-2.93 (m, 1H), 3.92 (d, J=1.53 Hz, 3H),4.06-4.11 (m, 2H), 4.51-4.57 (m, 2H), 5.07 (d, J=10.38 Hz, 1H), 5.25 (d,J=17.09 Hz, 1H), 5.78-5.85 (m, 2H), 7.10 (d, J=8.85 Hz, 1H), 7.17 (s,1H), 7.23 (d, J=4.27 Hz, 1H), 7.88 (dd, J=5.95, 1.68 Hz, 1H), 8.10 (d,J=9.16 Hz, 1H); MS m/z 726 (MH⁺).

Example 150 Preparation of Compound 150

Compound 150 was prepared by following Scheme 1 of Example 141 exceptthat the scale was increased and that L-valine benzyl esterhydrochloride was used in place of L-valine methyl ester hydrochloridein step 2.

Step 1:

Modifications: 100 mg (0.146 mmol) of the product of Example 55, Step 1;62.2 mg (0.219 mmol) N,N′-disuccinimidyl carbonate, 47.0 mg (0.364 mmol)N,N-diisopropylethylamine used.

Step 2:

Modifications: 356 mg (1.46 mmol) L-valine benzyl ester hydrochloride,188 mg (1.46 mmol) N,N-diisopropylethylamine used, 41.0 mg productobtained as a white powder (33.2% yield): MS m/z 848 (MH⁺).

Example 151 Preparation of Compound 151

Compound 151 was prepared by following Scheme 1 of Example 141 exceptthat the scale was increased and that L-isoleucine methyl esterhydrochloride was used in place of L-valine methyl ester hydrochloridein step 2.

Step 1:

Modifications: 100 mg (0.146 mmol) of the product of Example 55, Step 1;62.2 mg (0.219 mmol) N,N′-disuccinimidyl carbonate, 47.0 mg (0.364 mmol)N,N-diisopropylethylamine used.

Step 2:

Modifications: 265 mg (1.46 mmol) L-isoleucine methyl esterhydrochloride, 188 mg (1.46 mmol) N,N-diisopropylethylamine used, 75.5mg product obtained as a white powder (65.9% yield): ¹H NMR (CD₃OD) δ0.78-0.80 (m, 3H), 0.83-0.84 (m, 3H), 0.90-0.95 (m, 1H), 1.01-1.03 (m,2H), 1.06 (d, J=3.05 Hz, 9H), 1.17-1.21 (m, 2H), 1.32-1.42 (m, 2H),1.68-1.72 (m, 1H), 1.84-1.87 (m, 1H), 2.14-2.20 (m, 1H), 2.30-2.36 (m,1H), 2.57-2.62 (m, 1H), 2.90-2.95 (m, 1H), 3.66 (d, J=2.75 Hz, 3H), 3.92(d, J=2.75 Hz, 3H), 4.05-4.12 (m, 2H), 4.39-4.42 (m, 2H), 4.50-4.53 (m,1H), 5.07-5.10 (m, 1H), 5.23-5.28 (m, 1H), 5.73-5.79 (m, 1H), 5.81-5.83(m, 1H), 7.10-7.13 (m, 1H), 7.17 (t, J=2.44 Hz, 1H), 7.22-7.24 (m, 1H),7.85-7.87 (m, 1H), 8.10 (dd, J=9.16, 2.75 Hz, 1H); MS m/z 785 (MH⁺).

Example 152 Preparation of Compound 152

Compound 152 was prepared by following Scheme 1 of Example 141 exceptthat the scale was increased and that L-valine tert-butyl esterhydrochloride was used in place of L-valine methyl ester hydrochloridein step 2.

Step 1:

Modifications: 100 mg (0.146 mmol) of the product of Example 55, Step 1;62.2 mg (0.219 mmol) N,N′-disuccinimidyl carbonate, 47.0 mg (0.364 mmol)N,N-diisopropylethylamine used.

Step 2:

Modifications: 306 mg (1.46 mmol) L-valine tert-butyl esterhydrochloride, 188 mg (1.46 mmol) N,N-diisopropylethylamine used, 93.5mg product obtained as a white powder (78.8% yield): MS m/z 814 (MH⁺).

Example 153 Preparation of Compound 153

Compound 153 was prepared by following Scheme 1 of Example 141 exceptthat the scale was increased and that (S)-(+)-1-methoxy-2-propylaminewas used in place of L-valine methyl ester hydrochloride in step 2.

Step 1:

Modifications: 100 mg (0.146 mmol) of the product of Example 55, Step 1;62.2 mg (0.219 mmol) N,N′-disuccinimidyl carbonate, 47.0 mg (0.364 mmol)N,N-diisopropylethylamine used.

Step 2:

Modifications: 130 mg (1.46 mmol) (S)-(+)-1-methoxy-2-propylamine, 188mg (1.46 mmol) N,N-diisopropylethylamine used, 50.3 mg product obtainedas a white powder (47.3% yield): ¹H NMR (CD₃OD) δ 0.96 (d, J=7.02 Hz,3H), 0.99-1.01 (m, 2H), 1.04 (s, 9H), 1.15-1.18 (m, 2H), 1.39 (dd,J=9.61, 5.34 Hz, 1H), 1.84 (dd, J=7.93, 5.19 Hz, 1H), 1.93 (s, 3H), 2.15(q, J=9.05 Hz, 1H), 2.37 (ddd, J=13.96, 9.84, 4.58 Hz, 1H), 2.61 (dd,J=14.04, 7.32 Hz, 1H), 2.90 (ddd, J=12.89, 8.16, 4.88 Hz, 1H), 3.19-3.28(m, 2H), 3.64-3.67 (m, 1H), 3.92 (s, 3H), 4.12 (dd, J=11.60, 3.97 Hz,1H), 4.40 (s, 1H), 4.44 (d, J=11.90 Hz, 1H), 4.53 (dd, J=9.77, 7.32 Hz,1H), 5.07 (dd, J=10.22, 1.68 Hz, 1H), 5.24 (dd, J=17.24, 1.37 Hz, 1H),5.76-5.81 (m, 1H), 5.83 (s, 1H), 7.10 (dd, J=9.16, 2.44 Hz, 1H), 7.17(d, J=2.44 Hz, 1H), 7.23 (d, J=5.80 Hz, 1H), 7.87 (d, J=5.80 Hz, 1H),8.10 (d, J=9.16 Hz, 1H); MS m/z 729 (MH⁺).

Example 154 Preparation of Compound 154

Compound 154 was prepared by following Scheme 1 of Example 141 exceptthat the scale was increased and that N-methyl L-valine methyl esterhydrochloride was used in place of L-valine methyl ester hydrochloridein step 2.

Step 1:

Modifications: 100 mg (0.146 mmol) of the product of Example 55, Step 1;62.2 mg (0.219 mmol) N,N′-disuccinimidyl carbonate, 47.0 mg (0.364 mmol)N,N-diisopropylethylamine used.

Step 2:

Modifications: 265 mg (1.46 mmol) N-methyl L-valine methyl esterhydrochloride, 188 mg (1.46 mmol) N,N-diisopropylethylamine used, 68.2mg product obtained as a white powder (59.5% yield): ¹H NMR (CD₃OD) δ0.70 (dd, J=6.71, 2.14 Hz, 3H), 0.89 (dd, J=6.41, 2.44 Hz, 3H),0.96-0.98 (m, 1H), 1.02-1.04 (m, 2H), 1.07 (d, J=2.14 Hz, 9H), 1.18-1.22(m, 2H), 1.43-1.47 (m, 1H), 1.84-1.87 (m, 1H), 2.11-2.19 (m, 2H),2.31-2.37 (m, 1H), 2.58-2.63 (m, 1H), 2.87 (d, J=2.44 Hz, 3H), 2.90-2.94(m, 1H), 3.65 (d, J=2.14 Hz, 3H), 3.92 (d, J=2.14 Hz, 3H), 4.10-4.14 (m,1H), 4.25 (dd, J=10.07, 1.22 Hz, 1H), 4.48 (d, J=2.44 Hz, 1H), 4.50-4.54(m, 1H), 5.08-5.10 (m, 1H), 5.25-5.28 (dd, J=17.09, 1.53 Hz, 1H),5.78-5.85 (m, 2H), 7.10-7.13 (m, 1H), 7.18-7.19 (m, 1H), 7.24 (dd,J=5.95, 2.59 Hz, 1H), 7.87-7.89 (m, 1H), 8.10 (dd, J=9.00, 2.59 Hz, 1H);MS m/z 785 (MH⁺).

Example 155 Preparation of Compound 155

Step 1:

To a solution of the product of Example 55, Step 1 (100 mg, 0.146 mmol)in anhydrous THF (2 mL) was added carbonic acid pyridin-2-yl ester2,2,2-trifluoro-1,1-dimethyl-ethyl ester (44.0 mg, 0.175 mmol) andN-methylmorpholine (59 mg, 0.58 mmol). The mixture was agitated at rtfor 24 h. The reaction mixture was washed concentrated in vacuo and theresidue was dissolved in DCM (2 mL). The solution was washed with pH=4buffer solution (3×3 mL), and the washes were back-extracted with DCM (3mL). The organic phases were combined and concentrated in vacuo. Thecrude product was then dissolved in MeOH and purified by reverse phasepreparative HPLC to give Compound 155 as a white powder (38.5 mg, 34.3%yield): ¹H NMR (CD₃OD) δ 1.04 (s, 11H), 1.19-1.22 (m, 2H), 1.23 (s, 3H),1.43 (dd, J=9.31, 5.34 Hz, 1H), 1.46 (s, 3H), 1.87 (dd, J=7.93, 5.49 Hz,1H), 2.19 (q, J=8.85 Hz, 1H), 2.34 (m, 1H), 2.62 (dd, J=13.73, 7.02 Hz,1H), 2.92 (ddd, J=12.67, 8.09, 4.88 Hz, 1H), 3.92 (s, 3H), 4.06 (dd,J=11.90, 3.36 Hz, 1H), 4.23 (s, 1H), 4.43 (d, J=11.60 Hz, 1H), 4.56 (dd,J=10.38, 7.32 Hz, 1H), 5.10 (d, J=10.38 Hz, 1H), 5.27 (d, J=17.09 Hz,1H), 5.75-5.80 (m, 1H), 5.82 (s, 1H), 7.10 (dd, J=9.16, 2.44 Hz, 1H),7.18 (d, J=2.44 Hz, 1H), 7.25 (d, J=6.10 Hz, 1H), 7.89 (d, J=5.80 Hz,1H), 8.07 (d, J=9.16 Hz, 1H); MS m/z 768 (MH⁺).

Section E

LC-MS conditions for section E

“method A” is 3.0×50 mm Xterra 4 min gradient and 4 mL/min flow

“method B” is 3.0×50 mm Xterra 3 min gradient and 4 mL/min flow

“method C” is 4.6×50 mm Xterra 4 min gradient and 4 mL/min flow

“method D” is 4.6×50 mm Xterra 3 min gradient and 4 mL/min flow

Example 180 Preparation of Compound 180

Compound 180 to 183 were prepared by the general synthetic scheme asdepicted above. These individual reactions were described in detailelsewhere. With the exception of the first alkylation step for whichpotassium tert-butoxide in THF (as supplied by Aldrich Chemicals) in DMFoffered a more convenient work up procedure: most of the DMF solvent waswashed away with water once the alkylation was complete.

Thus Compound 180: BOCNH-P3(L-tert-BuGly)-P2[(4R)-(2-trifluoromethylquinolin-4-oxo)-S-proline]-P1(1R,2S Vinyl Acca)-CONHSO₂-Cyclopropane:the material was obtained as a white foam in 61% yield. LC/MS R_(t)-min(MNa⁺) [method A]: 3.35 (774). ¹H NMR (400 MHz, CD₃OD) δ ppm 1.05 (m,13H) 1.21 (s, 9H) 1.42 (dd, J=9.17, 5.26 Hz, 1H) 1.86 (dd, J=8.07, 5.38Hz, 1H) 2.21 (m, 1H) 2.33 (m, 1H) 2.64 (dd, J=13.94, 6.60 Hz, 1H) 2.93(m, 1H) 4.09 (dd, J=11.49, 2.69 Hz, 1H) 4.21 (s, 1H) 4.52 (m, 1H) 4.56(d, J=12.23 Hz, 1H) 5.10 (dd, J=10.39, 1.59 Hz, 1H) 5.27 (d, J=16.87 Hz,1H) 5.58 (s, 1H) 5.72 (m, 1H) 7.36 (s, 1H) 7.61 (t, J=7.70 Hz, 1H) 7.83(t, J=7.34 Hz, 1H) 8.07 (d, J=8.56 Hz, 1H) 8.26 (d, J=8.56 Hz, 1H).

Example 181 Preparation of Compound 181

BOCNH-P3(L-tert-BuGly)-P2[(4R)-(2,8-bistrifluoromethylquinolin-4-oxo)-S-proline]-P1(1R,2S Vinyl Acca)-CONHSO₂-Cyclopropane:the material was obtained as a white foam in 52% yield. LC/MS R_(t)-min(MNa⁺) [method A]: 3.60 (843). ¹H NMR (400 MHz, CD₃OD) δ ppm 1.05 (m,11H) 1.16 (s, 9H) 1.22 (m, 2H) 1.42 (dd, J=9.29, 5.38 Hz, 1H) 1.86 (dd,J=8.07, 5.38 Hz, 1H) 2.21 (q, J=8.97 Hz, 1H) 2.33 (m, 1H) 2.65 (dd,J=13.94, 6.85 Hz, 1H) 2.93 (m, 1H) 4.07 (dd, J=11.98, 2.69 Hz, 1H) 4.17(s, 1H) 4.52 (dd, J=10.52, 6.85 Hz, 1H) 4.58 (d, J=11.98 Hz, 1H) 5.10(d, J=10.27 Hz, 1H) 5.27 (d, J=17.12 Hz, 1H) 5.60 (s, 1H) 5.72 (m, 1H)7.46 (s, 1H) 7.69 (t, J=7.83 Hz, 1H) 8.18 (d, J=7.34 Hz, 1H) 8.50 (d,J=8.31 Hz, 1H).

Example 182 Preparation of Compound 182

BOCNH-P3(L-tert-BuGly)-P2[(4R)-(2-trifluoromethyl, 8-trifluoromethoxyquinolin-4-oxo)-S-proline]-P1(1R,2S Vinyl Acca)-CONHSO₂-Cyclopropane:the material was obtained as a white foam in 99% yield. LC/MS R_(t)-min(MNa⁺) [method A]: 3.62 (858). ¹H NMR (400 MHz, CD₃OD) δ ppm 1.05 (m,11H) 1.21 (m, 11H) 1.42 (dd, J=9.05, 5.14 Hz, 1H) 1.86 (dd, J=8.07, 5.38Hz, 1H) 2.21 (q, J=8.64 Hz, 1H) 2.33 (m, 1H) 2.64 (dd, J=13.94, 6.60 Hz,1H) 2.93 (m, 1H) 4.08 (dd, J=11.98, 2.69 Hz, 1H) 4.18 (s, 1H) 4.51 (dd,J=10.52, 6.85 Hz, 1H) 4.57 (d, J=12.23 Hz, 1H) 5.10 (dd, J=10.52, 1.22Hz, 1H) 5.27 (d, J=17.12 Hz, 1H) 5.59 (s, 1H) 5.72 (m, 1H) 7.44 (s, 1H)7.63 (t, J=8.07 Hz, 1H) 7.78 (d, J=7.58 Hz, 1H) 8.24 (d, J=8.56 Hz, 1H).

Example 183 Preparation of Compound 183

BOCNH-P3(L-tert-BuGly)-P2[(4R)-(2-trifluoromethyl, 8-chloroquinolin-4-oxo)-S-proline]-P1(1R,2S Vinyl Acca)-CONHSO₂-Cyclopropane:the material was obtained as a white foam in 64% yield. LC/MS R_(t)-min(MNa⁺) [method A]: 3.52 (808). ¹H NMR (400 MHz, CD₃OD) δ ppm 1.04 (m,11H) 1.21 (m, 11H) 1.41 (dd, J=9.41, 5.50 Hz, 1H) 1.86 (dd, J=8.07, 5.62Hz, 1H) 2.20 (q, J=8.80 Hz, 1H) 2.32 (m, 1H) 2.63 (dd, J=13.82, 6.72 Hz,1H) 2.92 (m, 1H) 4.07 (dd, J=12.10, 2.81 Hz, 1H) 4.19 (s, 1H) 4.50 (dd,J=10.52, 6.85 Hz, 1H) 4.56 (d, J=11.98 Hz, 1H) 5.10 (dd, J=10.27, 1.47Hz, 1H) 5.26 (d, J=17.12 Hz, 1H) 5.57 (s, 1H) 5.72 (m, 1H) 7.41 (s, 1H)7.52 (t, J=8.07 Hz, 1H) 7.93 (d, J=7.58 Hz, 1H) 8.19 (d, J=8.56 Hz, 1H).

Example 184 General Procedure for Alkylation with the Tripeptide(Compound 184) and P2*

General Scheme—Preparation of Example 184 (Compound 184)

The preparation of the tripeptide component, Example 184 was achieved bya sequential amide coupling using HATU as the coupling agent. It isunderstood that many standard coupling agents could be employed for thefollowing scheme.

Preparation of intermediate Example 184a

To a mixture of HATU (820 mg, 2.2 mmol), Example 180a(Boc-4R-hydroxyproline, 417 mg, 1.8 mmol) and Example 180c(cyclopropanesulfonic acid(1(R)-amino-2(S)-vinyl-cyclopropanecarbonyl)-amide hydrochloride salt,490 mg, 1.8 mmol) in a flame dried flask at room temperature was addeddry CH₂Cl₂ (8 mL). The mixture was kept under dry N₂ before it waschilled to −78° C. Hunig's base (diisopropylethylamine, 625 μL, 3.6mmol) was added slowly over a period 5 min and the mixture turned into apale orange suspension. Stirring was continued for an hour whiletemperature was allow to raise to ambient. LC/MS showed completeconversion into the desired product 184a. The crude reaction was workedup as usual, washed with three portions (5 mL) of water, organicresidues were extracted into ethyl acetate (3×5 mL). The crude productwas obtained by removal of organic solvents in vacuo. The material wasused in the next step without further purification.

Preparation of intermediate Example 184b

The dried solid from the previous step was taken into 9 mL of CH₂Cl₂ atroom temperature. To this solution was added 3 mL of trifluoroaceticacid forming a pale yellowish solution. Stirring was continued for 2hours at room temperature. LC/MS showed no starting material 184a whilethe desired product 184b was the major signal along with a signalcorresponding to a side product carried over from HATU in the stepearlier. The solvents were evaporated and the solid residue was used inthe next step immediately without further purification.

Preparation of the Tripeptide, Compound 184

BOCNH-P3(L-tert-BuGly)-P2[(4R)-hydroxy-5-proline]-P1(1R,2S VinylAcca)-CONHSO₂-Cyclopropane [Notebook 46877-128]

The crude product from the previous step (Example 184b, 1.8 mmol) wasmixed with HATU (700 mg, 1.8 mmol) and BOC-L-tert-leucine (Example 180f,Fluka Chemicals, 420 mg, 1.8 mmol) in CH₂Cl₂ (10 mL) at roomtemperature. To this suspension was treated Hunig's base (1 mL, excess)forming a somewhat thinner, orange suspension. LC/MS showed someconversion into compound 184. Complete conversion into the desiredproduct 184 was observed after stirring was continued for two days atroom temperature. Crude reaction mixture was evaporated to dryness. Theresidue was taken into ethyl acetate. Most of HATU residue was removedby extractions with half saturated, freshly prepared sodium bicarbonatesolution. The last traces of HATU residue (1-hydroxy 7-azabenzotriazole)was removed by washing with deionized water. Evaporation of solventsgave 990 mg (98%) of the desired product as white foam. This material issuitable for the subsequent alkylation with electrophiles such asquinolines and isoquinolines directly without further purification.LC/MS R_(t)-min (MNa⁺) [method A]: 2.65 (579). ¹H NMR (400 MHz, CD₃OD) δppm 0.95 (s, 2H) 0.99 (s, 9H) 1.05 (m, 2H) 1.21 (m, 1H) 1.39 (m, 9H)1.84 (dd, J=8.31, 5.38 Hz, 1H) 1.96 (m, 1H) 2.11 (m, 1H) 2.20 (m, 1H)2.91 (m, 1H) 3.80 (m, 2H) 4.28 (d, J=9.78 Hz, 1H) 4.35 (dd, J=9.90, 6.97Hz, 1H) 4.47 (s, 1H) 5.11 (m, 1H) 5.29 (d, J=17.12 Hz, 1H) 5.75 (m, 1H).

Alkylation of the tripeptide (compound 184) with electrophiles:

To a flame-dried 25 mL round bottom flask was charged with Compound 184,(0.5-1.0 mmol), substituted 4-chloroquinoline (1.0 equivalent) andlanthanum chloride (LaCl₃ anhydrous beads, used as supplied by Aldrich,M.W. 245 g/mol; 1.0 equivalent. Note: the inclusion of such additive wasfound to be helpful in some cases especially with those less reactiveelectrophiles. This reagent can, at times, be omitted if theelectrophiles are sufficiently reactive towards anionic alkylation) in 2mL dry DMF. The inorganic salt was only sparingly soluble in DMF at roomtemperature. The mixture was chilled to −78° C. (dry-ice/acetone bath)with stirring under nitrogen. To this chilled mixture was added a THFsolution of potassium tert-butoxide (1.0 M, used as supplied by Aldrich,5.5 equivalents) and the color of mixture changed from colorless to paleyellowish or greenish. It was stirred at −78° C. for a period dependentupon the 4-chloroquinoline reactivity (a few hrs. at −78° C. toovernight at room temperature). The inorganic salt was also found tochange into a fine emulsion at the end. It was quenched with a halfsaturated NH₄Cl aqueous solution (2 mL). Organic materials wereextracted into ethyl acetate (10 mL×3). Organic layers were combined,back washed with deionized water (10 mL×2). Evaporation of the organicfraction gave a crude mixture rich in the desired product as determinedby LC/MS. The desired product was isolated by preparative HPLC usingstandard separation parameters (typically: 3.0×50 mm Xterra column 4 mingradient and 4 mL/min flow rate) to give the analytically pure desiredproduct. The alkylation of 1-halo isoquinoline series was carried out inexactly the same way.

Example 185 Preparation of Compound 185

Following the general tripeptide alkylation procedure as described inExample 184, BOCNH-P3(L-tert-BuGly)-P2[(4R)-(7-trifluoromethylquinolin-4-oxo)-S-proline]-P1(1R,2S Vinyl Acca)-CONHSO₂-Cyclopropane wasobtained as a white foam in 50% yield. LC/MS R_(t)-min (MH⁺) [method B]:2.32 (752). ¹H NMR (400 MHz, CD₃OD) δ ppm 1.02 (s, 9H) 1.06 (m, 11H)1.22 (m, 2H) 1.43 (dd, J=9.41, 5.26 Hz, 1H) 1.88 (dd, J=8.19, 5.50 Hz,1H) 2.23 (q, J=8.80 Hz, 1H) 2.42 (m, 1H) 2.75 (dd, J=14.06, 6.48 Hz, 1H)2.93 (m, 1H) 4.10 (m, 2H) 4.61 (m, 2H) 5.12 (dd, J=10.39, 1.59 Hz, 1H)5.29 (d, J=17.12 Hz, 1H) 5.72 (m, 2H) 7.61 (d, J=6.36 Hz, 1H) 7.96 (d,J=8.80 Hz, 1H) 8.38 (s, 1H) 8.59 (d, J=8.56 Hz, 1H) 9.14 (d, J=6.36 Hz,1H).

Example 186 Preparation of Compound 186

Following the general tripeptide alkylation procedure as described inExample 184, BOCNH-P3(L-tert-BuGly)-P2[(4R)-(8-trifluoromethylquinolin-4-oxo)-S-proline]-P1(1R,2S Vinyl Acca)-CONHSO₂-Cyclopropane:the desired product was obtained as a white foam in 50% yield. LC/MSR_(t)-min (MH⁺) [method B]: 2.48 (752). ¹H NMR (400 MHz, CD₃OD) δ ppm1.02 (s, 9H) 1.05 (m, 2H) 1.13 (s, 9H) 1.23 (m, 2H) 1.42 (dd, J=8.68,5.50 Hz, 1H) 1.87 (dd, J=8.07, 5.38 Hz, 1H) 2.21 (q, J=8.80 Hz, 1H) 2.36(m, 1H) 2.69 (dd, J=14.06, 6.97 Hz, 1H) 2.93 (m, 1H) 4.08 (dd, J=11.98,2.93 Hz, 1H) 4.15 (s, 1H) 4.54 (dd, J=10.52, 7.09 Hz, 1H) 4.60 (d,J=12.47 Hz, 1H) 5.11 (dd, J=10.52, 1.71 Hz, 1H) 5.28 (d, J=15.90 Hz, 1H)5.58 (s, 1H) 5.72 (m, 1H) 7.32 (d, J=5.87 Hz, 1H) 7.69 (t, J=7.95 Hz,1H) 8.22 (d, J=7.09 Hz, 1H) 8.55 (d, J=8.07 Hz, 1H) 8.88 (d, J=5.62 Hz,1H).

Preparation of isoquinoline intermediates for 6-F, 6-Ethyl, 6-isopropyland 6-tert-butyl isoquinoline P2* building blocks.

In general, the 6-fluoro and 6-alkyl isoquinolines used in the followingexperiments were prepared via a Pomeranz-Fritsch synthesis (Typicalprocedure: Preparation of optically active 8,8-disubstituted1,1-biisoquinoline, K. Hirao, R. Tsuchiya, Y. Yano, H. Tsue,Heterocycles 42(1) 1996, 415-422) as outlined below. The products wereconverted into the 1-chloro derivatives via N-oxide intermediates asdescribed elsewhere.

Reagents and reaction conditions: (a) reflux in benzene, azeotropicremoval of water; (b) first step: ethyl chloroformate, trimethylphosphite in THF, second step: titanium tetrachloride in chloroform; (c)MCPBA in CH₂Cl₂; (d) POCl₃ in benzene

R Isoquinoline, Yield 1-Chloride, combined yield F 20 43 Et 76 65 i-Pr14 18 t-Bu 47 55

Example 187 Preparation of Compound 187

BOCNH-P3(L-tert-BuGly)-P2[(4R)-(6-fluoroisoquinolin-1-oxo)-S-proline]-P1(1R,2S Vinyl Acca)-CONHSO₂-Cyclopropane:the material was obtained as a white foam in 12% yield. LC/MS R_(t)-min(MNa⁺) [method C]: 3.81 (724). ¹H NMR (400 MHz, CD₃OD) δ ppm 1.05 (m,13H) 1.22 (s, 9H) 1.42 (m, 1H) 1.86 (m, 1H) 2.21 (m, 2H) 2.61 (dd,J=13.69, 6.60 Hz, 1H) 2.93 (m, 1H) 4.05 (d, J=13.69 Hz, 1H) 4.21 (s, 1H)4.49 (m, 2H) 5.11 (d, J=10.03 Hz, 1H) 5.28 (d, J=17.61 Hz, 1H) 5.72 (m,1H) 5.86 (d, J=4.40 Hz, 1H) 7.31 (m, 2H) 7.48 (d, J=8.31 Hz, 1H) 7.97(d, J=6.36 Hz, 1H) 8.26 (d, J=6.11 Hz, 1H).

Example 188 Preparation of Compound 188

The alkylation described above gave the 1-chloroisoquinoline as themajor product: BOCNH-P3(L-tert-BuGly)-P2[(4R)-(1-chloroisoquinolin-6-oxo)-S-proline]-P1(1R,2S Vinyl Acca)-CONHSO₂-Cyclopropane:the material was obtained as a white foam in 40.2% yield. LC/MSR_(t)-min (MH⁺) [method C]: 3.81 (718). ¹H NMR (400 MHz, CD₃OD) δ ppm1.00 (s, 9H) 1.06 (m, 2H) 1.25 (s, 11H) 1.41 (m, 1H) 1.86 (dd, J=8.07,5.38 Hz, 1H) 2.25 (m, 2H) 2.54 (dd, J=12.96, 6.60 Hz, 1H) 2.92 (m, 1H)4.06 (dd, J=11.98, 2.69 Hz, 1H) 4.20 (s, 1H) 4.31 (d, J=11.74 Hz, 1H)4.45 (dd, J=9.78, 7.58 Hz, 1H) 5.11 (dd, J=10.27, 1.71 Hz, 1H) 5.28 (dd,J=17.36, 1.47 Hz, 1H) 5.36 (s, 1H) 5.74 (m, 1H) 7.35 (d, J=9.29 Hz, 1H)7.40 (s, 1H) 7.70 (d, J=5.87 Hz, 1H) 8.13 (d, J=5.87 Hz, 1H) 8.25 (d,J=9.29 Hz, 1H).

Example 189 Preparation of Compound 189

BOCNH-P3(L-tert-BuGly)-P2[(4R)-(6-ethylisoquinolin-1-oxo)-S-proline]-P1(1R,2S Vinyl Acca)-CONHSO₂-Cyclopropane:the material was obtained as a white foam in 4.6 mg of yellow solid wasobtained (4.2%). LC/MS rt-min (MH⁺) [method B]: 2.70 (712). ¹H NMR (400MHz, CD₃OD) δ ppm 1.01 (s, 9H) 1.07 (m, 2H) 1.22 (m, 11H) 1.28 (t,J=7.91 Hz, 3H) 1.41 (m, 1H) 1.85 (m, 1H) 2.25 (m, 2H) 2.60 (dd, J=13.69,6.85 Hz, 1H) 2.80 (q, J=7.66 Hz, 2H) 2.93 (m, 1H) 4.02 (d, J=31.06 Hz,1H) 4.23 (s, 1H) 4.42 (m, 1H) 4.54 (m, 1H) 5.10 (d, J=10.27 Hz, 1H) 5.28(d, J=17.12 Hz, 1H) 5.74 (m, 1H) 5.84 (s, 1H) 7.25 (d, J=5.87 Hz, 1H)7.38 (d, J=8.56 Hz, 1H) 7.59 (s, 1H) 7.90 (d, J=6.24 Hz, 1H) 8.09 (d,J=8.56 Hz, 1H).

Example 190 Preparation of Compound 190

BOCNH-P3(L-tert-BuGly)-P2[(4R)-(6-isopropylisoquinolin-1-oxo)-S-proline]-P1(1R,2S Vinyl Acca)-CONHSO₂-Cyclopropane:the material was obtained as a white foam in 69% yield. LC/MS R_(t)-min(MNa⁺) [method B]: 2.76 (749). ¹H NMR (400 MHz, CD₃OD) δ ppm 1.05 (m,13H) 1.20 (m, 9H) 1.31 (d, J=6.85 Hz, 6H) 1.42 (m, 1H) 1.86 (dd, J=8.07,5.62 Hz, 1H) 2.26 (m, 2H) 2.62 (dd, J=13.69, 6.85 Hz, 1H) 2.93 (m, 1H)3.07 (m, 1H) 4.06 (m, 1H) 4.21 (s, 1H) 4.52 (m, 2H) 5.11 (d, J=10.27 Hz,1H) 5.28 (d, J=17.12 Hz, 1H) 5.74 (m, 1H) 5.84 (s, 1H) 7.32 (d, J=6.11Hz, 1H) 7.46 (d, J=8.56 Hz, 1H) 7.64 (s, 1H) 7.90 (d, J=6.11 Hz, 1H)8.13 (d, J=8.56 Hz, 1H).

Example 191 Preparation of Compound 191

BOCNH-P3(L-tert-BuGly)-P2[(4R)-(6-tert-butylisoquinolin-1-oxo)-S-proline]-P1(1R,2S Vinyl Acca)-CONHSO₂-Cyclopropane:the material was obtained as a white foam in 81% yield. LC/MS R_(t)-min(MH⁺) [method B]: 2.84 (740). ¹H NMR (400 MHz, CD₃OD) δ ppm 1.01 (s, 9H)1.06 (m, 2H) 1.18 (s, 9H) 1.22 (m, 2H) 1.39 (s, 9H) 1.43 (m, 1H) 1.87(dd, J=8.19, 5.50 Hz, 1H) 2.27 (m, 2H) 2.63 (dd, J=13.57, 6.97 Hz, 1H)2.93 (m, 1H) 4.06 (m, 1H) 4.20 (s, 1H) 4.52 (m, 2H) 5.10 (d, J=11.49 Hz,1H) 5.28 (d, J=17.12 Hz, 1H) 5.73 (m, 1H) 5.84 (s, 1H) 7.36 (d, J=6.11Hz, 1H) 7.66 (dd, J=8.80, 1.22 Hz, 1H) 7.78 (s, 1H) 7.91 (d, J=5.87 Hz,1H) 8.15 (d, J=8.80 Hz, 1H).

Preparation of 6-isopropoxyl and 6-tert-butoxyl isoquinolineIntermediates

Some 6-alkoxy-1-chloro isoquinolines were prepared by a direct, ipsodisplacement of the 6-fluoro-1-chloroisoquinoline with the correspondingalkoxide metal ions such as potassium tert-butoxide (53%) and sodiumisopropoxide (54%).

General Synthetic Scheme

The 6-fluoro-1-chloroisoquinoline was subjected to an aromaticnucleophilic displacement with sodium isopropoxide and potassiumtert-butoxide in DMF to give the corresponding

6-isopropoxyl (54%): ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 1.43 (d,J=6.11 Hz, 6H) 4.76 (m, J=6.11 Hz, 1H) 7.08 (d, J=2.45 Hz, 1H) 7.29 (dd,J=9.29, 2.45 Hz, 1H) 7.50 (d, J=5.62 Hz, 1H) 8.18 (d, J=5.87 Hz, 1H)8.24 (d, J=9.29 Hz, 1H) and 6-tert-butoxyl-1-chloro isoquinolines (55%):¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 1.48 (s, 9H) 7.31 (m, 2H) 7.47 (d,J=5.62 Hz, 1H) 8.18 (d, J=5.62 Hz, 1H) 8.21 (d, J=9.78 Hz, 1H) as themajor product respectively. These 6-alkoxyl-1-chloro isoquinolines werealkylated with the tripeptide as described in Example 184 to give thedesired products shown below.

Example 192 Preparation of Compound 192

BOCNH-P3(L-tert-BuGly)-P2[(4R)-(6-isopropoxyisoquinolin-1-oxo)-S-proline]-P1(1R,2S Vinyl Acca)-CONHSO₂-Cyclopropane:the material was obtained as a white foam in 59% yield. LC/MS R_(t)-min(MH⁺) [method C]: 3.87 (742). ¹H NMR (400 MHz, CD₃OD) δ ppm 0.99 (s,11H) 1.06 (m, 2H) 1.21 (s, 9H) 1.37 (d, J=5.87 Hz, 6H) 1.42 (m, 1H) 1.86(dd, J=8.07, 5.38 Hz, 1H) 2.25 (m, 2H) 2.61 (dd, J=13.69, 6.85 Hz, 1H)2.92 (m, 1H) 4.04 (dd, J=11.86, 3.30 Hz, 1H) 4.21 (br. s, 1H) 4.49 (m,2H) 4.78 (h, J=5.87 Hz, 1H) 5.10 (dd, J=10.39, 1.35 Hz, 1H) 5.28 (d,J=16.87 Hz, 1H) 5.73 (m, 1H) 5.79 (d, J=11.25 Hz, 1H) 7.07 (dd, J=9.05,1.96 Hz, 1H) 7.18 (d, J=2.20 Hz, 1H) 7.26 (d, J=6.11 Hz, 1H) 7.85 (d,J=6.11 Hz, 1H) 8.10 (d, J=9.05 Hz, 1H).

Example 193 Preparation of Compound 193

BOCNH-P3(L-tert-BuGly)-P2[(4R)-(6-tert-butoxyisoquinolin-1-oxo)-S-proline]-P1(1R,2S Vinyl Acca)-CONHSO₂-Cyclopropane:the material was obtained as a white foam in 39% yield. LC/MS R_(t)-min(MH⁺) [method C]: 3.99 (756). ¹H NMR (400 MHz, CD₃OD) δ ppm 1.03 (br s,13H) 1.19 (s, 9H) 1.42 (m, 10H) 1.86 (dd, J=7.83, 5.62 Hz, 1H) 2.24 (m,2H) 2.60 (dd, J=13.69, 6.85 Hz, 1H) 2.93 (m, 1H) 4.04 (dd, J=11.49, 2.93Hz, 1H) 4.23 (s, 1H) 4.49 (m, 2H) 5.10 (d, J=11.25 Hz, 1H) 5.27 (d,J=16.87 Hz, 1H) 5.73 (m, 1H) 5.81 (s, 1H) 7.13 (dd, J=8.80, 1.47 Hz, 1H)7.25 (d, J=5.87 Hz, 1H) 7.33 (d, J=2.20 Hz, 1H) 7.87 (d, J=6.11 Hz, 1H)8.10 (d, J=9.05 Hz, 1H).

Preparation of Phthalazine P2* Derivatives

In general, both 1-chlorophthalazine and 1,4-dichlorophthalazine undergoalkylation smoothly to give the desired products. However, thecommercially available 1-chlorophthalazine and 1,4-dichlorophthalazineare often contaminated with some hydrolyzed materials. A pre-treatmentwith POCl₃ followed by alkylation immediately afterward furnished moreconsistent results.

Reaction Conditions: (a) POCl₃ in DCE; (b) Alkylation with tripeptide;(c) sodium derivatives of imidazole (R═CH), triazole (R═N)

Example 194 Preparation Compound 194

BOCNH-P3(L-tert-BuGly)-P2[(4R)-(phthalazine-1-oxo)-S-proline]-P1(1R,2SVinyl Acca)-CONHSO₂-Cyclopropane: the material was obtained as a whitefoam in 41% yield. LC/MS R_(t)-min (MNa⁺) [method B]: 2.07 (707). ¹H NMR(400 MHz, CD₃OD) δ ppm 1.03 (m, 9H) 1.06 (m, 4H) 1.14 (s, 9H) 1.20 (m,1H) 1.43 (m, 1H) 1.87 (dd, J=8.07, 5.62 Hz, 1H) 2.24 (q, J=8.80 Hz, 1H)2.38 (m, 1H) 2.76 (dd, J=14.18, 7.09 Hz, 1H) 2.92 (m, 1H) 4.11 (m, 2H)4.62 (m, 1H) 5.11 (dd, J=10.27, 1.71 Hz, 1H) 5.29 (dd, J=17.12, 1.22 Hz,1H) 5.72 (m, 1H) 5.96 (s, 1H) 8.26 (m, 2H) 8.46 (m, 2H) 9.84 (s, 1H).

Example 195 Preparation of Compound 195

BOCNH-P3(L-tert-BuGly)-P2[(4R)-(4-chlorophthalazine-1-oxo)-S-proline]-P1(1R,2S Vinyl Acca)-CONHSO₂-Cyclopropane:the material was obtained as a white foam in 23% yield. LC/MS R_(t)-min(MNa⁺) [method C]: 3.52 (742). ¹H NMR (400 MHz, CD₃OD) δ ppm 1.01 (s,11H) 1.06 (m, 2H) 1.14 (s, 9H) 1.22 (m, 1H) 1.43 (m, 1H) 1.87 (m, 1H)2.21 (m, 1H) 2.35 (m, J=10.27 Hz, 1H) 2.70 (m, 1H) 2.93 (m, 1H) 4.05 (d,J=3.42 Hz, 1H) 4.58 (m, 2H) 5.11 (dd, J=10.39, 1.10 Hz, 1H) 5.28 (d,J=17.36 Hz, 1H) 5.73 (m, 1H) 5.93 (s, 1H) 7.99 (m, 1H) 8.07 (t, J=7.70Hz, 1H) 8.26 (dd, J=8.19, 2.32 Hz, 2H).

Preparation of 4-(imidazo-1-yl)phthalazine and4-(1,2,4-triazo-1-yl)phthalazine P2* Derivatives

The product, compound 195 from above,BOCNH-P3(L-tert-BuGly)-P2[(4R)-(4-chlorophthalazine-1-oxo)-S-proline]-P1(1R,2S Vinyl Acca)-CONHSO₂-Cyclopropane,was subjected to displacement by the anions of typical azoles such asimidazole and triazole to give 4-azole substituted phthalazinederivatives shown below:

Example 196 Preparation of Compound 196

This was made by displacing the 4-chloro phthalazine (Compound 195) withthe sodium salt of imidazole in DMF at 55-65° C.

BOCNH-P3(L-tert-BuGly)-P2[(4R)-(4-(imidazo-1-yl)phthalazine-1-oxo)-S-proline]-P1(1R,2SVinyl Acca)-CONHSO₂-Cyclopropane: the material was obtained as a whitefoam in 23% yield. LC/MS R_(t)-min (MNa⁺) [method B]: 1.94 (773). ¹H NMR(400 MHz, CD₃OD) δ ppm 1.03 (s, 9H) 1.07 (m, 2H) 1.20 (m, 9H) 1.24 (m,1H) 1.41 (m, 2H) 1.88 (dd, J=8.19, 5.50 Hz, 1H) 2.23 (m, 1H) 2.41 (m,1H) 2.75 (m, J=14.92 Hz, 1H) 2.93 (m, 1H) 4.14 (m, 2H) 4.60 (dd,J=10.15, 6.97 Hz, 2H) 5.12 (dd, J=10.27, 1.47 Hz, 1H) 5.29 (dd, J=17.24,1.35 Hz, 1H) 5.74 (m, 1H) 6.09 (s, 1H) 7.85 (s, 1H) 7.94 (m, 1H) 8.10(m, 2H) 8.43 (m, 1H) 9.17 (s, 1H) 9.46 (s, 1H).

During the displacement reaction, small amount of de-BOC by-product(Compound 197) was also isolated:

NH₂—P3(L-tert-BuGly)-P2[(4R)-(4-(imidazo-1-yl)phthalazine-1-oxo)-S-proline]-P1(1R,2SVinyl Acca)-CONHSO₂-Cyclopropane: the material was obtained as a whitefoam in 17% yield. LC/MS R_(t)-min (MH⁺) [method B]: 1.22 (651). ¹H NMR(400 MHz, CD₃OD) δ ppm 1.16 (s, 9H) 1.23 (m, 2H) 1.42 (dd, J=9.41, 5.50Hz, 1H) 1.89 (m, 1H) 2.26 (q, J=8.97 Hz, 1H) 2.43 (m, 1H) 2.78 (dd,J=14.06, 7.21 Hz, 1H) 2.93 (m, 1H) 3.74 (m, 1H) 4.11 (s, 1H) 4.22 (dd,J=12.23, 3.91 Hz, 1H) 4.47 (m, 2H) 4.71 (dd, J=10.27, 7.09 Hz, 1H) 5.12(m, 1H) 5.29 (d, J=17.36 Hz, 1H) 5.71 (m, 1H) 6.13 (t, J=3.67 Hz, 1H)7.87 (s, 1H) 7.97 (m, 1H) 8.14 (m, 3H) 8.41 (m, 1H) 9.47 (s, 1H).

Example 198 Preparation Compound 198

This was made by displacing the 4-chloro phthalazine (Compound 195) withthe sodium salt of 1,2,4-triazole in DMF at 55-65° C.

BOCNH-P3(L-tert-BuGly)-P2[(4R)-(4-(1,2,4-triazo-1-yl)phthalazine-1-oxo)-S-proline]-P1(1R,2SVinyl Acca)-CONHSO₂-Cyclopropane: the material was obtained as a whitefoam in 62% yield. LC/MS R_(t)-min (MNa⁺) [method C]: 3.35 (774). ¹H NMR(500 MHz, CD₃OD) δ ppm 0.97 (s, 9H) 1.01 (m, 2H) 1.10 (s, 9H) 1.16 (m,1H) 1.37 (m, 2H) 1.82 (m, 1H) 2.18 (d, J=8.55 Hz, 1H) 2.32 (m, 1H) 2.69(dd, J=13.58, 6.87 Hz, 1H) 2.88 (br s, 1H) 4.06 (d, J=11.60 Hz, 1H) 4.13(s, 1H) 4.53 (m, J=9.16 Hz, 1H) 4.61 (d, J=11.90 Hz, 1H) 5.06 (d,J=10.07 Hz, 1H) 5.23 (d, J=17.40 Hz, 1H) 5.68 (m, 1H) 5.98 (s, 1H) 7.97(m, 2H) 8.30 (m, J=11.90 Hz, 2H) 8.44 (d, J=7.63 Hz, 1H) 9.14 (s, 1H).

Preparation of 4-hydroxy and 4-alkoxy Phthalazine P2* Derivatives

Reaction Conditions: (a) sodium alkoxides such as methoxide, ethoxideand isopropoxide

Example 199 Preparation of Compound 199

The 4-chloro phthalazine (Example 195) was dissolved in dry isopropylalcohol at suspension was brought to reflux. The desired product, 4.5 mgof yellow solid was obtained (20.0%). LC/MS rt-min (MH⁺): 2.68 (743)[method B]. ¹H NMR (400 MHz, CD₃OD) δ ppm 1.02 (m, 11H) 1.20 (m, 11H)1.42 (m, 1H) 1.49 (d, J=6 Hz, 6H) 1.87 (dd, J=7.95, 5.50 Hz, 1H) 2.21(m, 1H) 2.23 (m, 1H) 2.66 (m, 1H) 2.93 (m, 1H) 4.08 (q, J=7.09 Hz, 1H)4.19 (s, 1H) 4.55 (m, 2H) 5.11 (d, J=10.27 Hz, 1H) 5.28 (d, J=17.61 Hz,1H) 5.48 (m, 1H) 5.70 (d, J=10.03 Hz, 1H) 5.81 (m, 1H) 7.90 (m, 2H) 8.16(m, 2H).

Example 200 Preparation of Compound 200

Likewise, the 4-ethoxy derivative was prepared:BOCNH-P3(L-t-BuGly)-P2[(4R)-(4-ethoxyphthalazine-1-oxo)-S-proline]-P1(1R,2SVinylAcca)-CONHSO₂ Cyclopropane.

4.0 mg of yellow solid was obtained (16.0%). LC/MS rt-min (MH⁺): 2.52(729) [method B]. ¹H NMR (400 MHz, CD₃OD) δ ppm 1.01 (s, 9H) 1.06 (m,2H) 1.16 (s, 9H) 1.24 (m, 2H) 1.43 (dd, J=9.78, 5.14 Hz, 1H) 1.53 (t,J=6.97 Hz, 3H) 1.87 (dd, J=8.19, 5.50 Hz, 1H) 2.22 (q, J=8.97 Hz, 1H)2.32 (m, 1H) 2.67 (m, 1H) 2.93 (m, 1H) 4.06 (d, J=8.56 Hz, 1H) 4.19 (s,1H) 4.56 (m, 4H) 5.10 (m, 1H) 5.30 (m, 1H) 5.74 (m, 1H) 5.82 (s, 1H)7.94 (m, 2H) 8.16 (d, J=7.83 Hz, 1H) 8.21 (m, 1H).

Example 201 Preparation of Compound 201

BOCNH-P3(L-t-BuGly)-P2[(4R)-(4-methoxyphthalazine-1-oxo)-S-proline]-P1(1R,2SVinylAcca)-CONHSO₂ Cyclopropane was prepared in 30.2% yield. LC/MSrt-min (MH⁺): 2.42 (715) [method B] ¹H NMR (400 MHz, CD₃OD) δ ppm 0.96(s, 9H) 1.07 (m, 2H) 1.20 (m, 11H) 1.43 (dd, J=9.29, 5.38 Hz, 1H) 1.87(dd, J=8.07, 5.62 Hz, 1H) 2.22 (q, J=8.80 Hz, 1H) 2.31 (m, 1H) 2.66 (d,J=8.07 Hz, 1H) 2.93 (m, 1H) 4.06 (dd, J=11.98, 3.18 Hz, 1H) 4.19 (d,J=3.42 Hz, 4H) 4.54 (m, 2H) 5.11 (m, 1H) 5.28 (d, J=17.36 Hz, 1H) 5.73(m, 1H) 5.83 (s, 1H) 7.95 (m, 2H) 8.19 (m, 2H).

Example 202 Preparation of Compound 202

An attempt was made to displace BOCNH-P3(L-tert-BuGly)-P2[(4R)-(4-chlorophthalazine-1-oxo)-S-proline]-P1(1R,2S Vinyl Acca)-CONHSO₂-Cyclopropane(Example 195) with the sodium salt of tetrazole gave mostly the4-hydroxy, hydrolyzed material.

BOCNH-P3(L-t-BuGly)-P2[(4R)-(4-hydroxyphthalazine-1-oxo)-S-proline]P1(1R,2SVinylAcca)-CONHSO₂ Cyclopropane was obtained (44.2%) as a pale creamysolid. LC/MS rt-min (MH⁺): 2.18 (701) [method B]. ¹H NMR (400 MHz,CD₃OD) δ ppm 1.01 (s, 9H) 1.05 (m, 2H) 1.23 (m, 11H) 1.42 (dd, J=9.29,5.38 Hz, 1H) 1.87 (dd, J=8.07, 5.38 Hz, 1H) 2.21 (m, 2H) 2.63 (m, 1H)2.93 (m, 1H) 4.00 (s, 1H) 4.20 (s, 1H) 4.50 (m, 2H) 5.11 (dd, J=10.27,1.47 Hz, 1H) 5.29 (d, J=16.87 Hz, 1H) 5.59 (s, 1H) 5.73 (m, 1H) 7.86(dd, J=5.75, 3.30 Hz, 2H) 8.01 (dd, J=5.87, 3.42 Hz, 1H) 8.29 (dd,J=5.87, 3.42 Hz, 1H).

Preparation of 5,6-Disubstituted Isoquinoline P2* Derivatives Via anAlkylation Protocol

Example 203 Preparation of 1-chloro-5-propylthio-6-fluoro isoquinoline

To a chilled (˜78° C.) solution of 1-chloro-6-fluoro isoquinoline (59mg, 0.32 mmol) in 2 mL of THF was added LDA solution in cyclohexane (1.5Molar, 0.23 mL, 0.35 mmol). The orange solution was stirred for 2 hrsbefore it was treated with n-propyl disulfide (60 μL, neat material,excess). The reaction was allowed to warm to room temperature over 30min. It was quenched with a solution of half saturated NH₄Cl, theorganic residues were extracted into ethyl acetate. LC-MS analysisindicated about 50% conversion into the desired product along withmainly starting material.

The desired product was purified by a short column (4 cm×2 cm, silicagel type-H) eluted with 5% ether in hexanes, 29 mg (36% yield) of thedesired product was obtained. LC/MS R_(t)-min (MH⁺) [method C]: 3.79(256). ¹H NMR (400 MHz, CHLOROFORM-D) δ ppm 0.96 (t, J=7.34 Hz, 3H) 1.52(m, 2H) 2.86 (m, 2H) 7.45 (dd, J=9.29, 8.56 Hz, 1H) 8.34 (d, J=0.73 Hz,2H) 8.37 (m, 1H). This compound was alkylated with the tripeptide by wayof the procedure described in Example 184 to give the followingcompound:

Example 204 Preparation of Compound 204

BOCNH-P3(L-t-BuGly)-P2[(4R)-(1-Chloro-5-propylthio-isoquinolin-6-oxo)-S-proline]-P1(1R,2SVinylAcca)-CONHSO₂ Cyclopropane, Shown below

Following the general procedure, 4.6 mg of yellow solid was obtained(3.2%). LC/MS rt-min (MH⁺): 2.73 (792) (method B). ¹H NMR (400 MHz,CD₃OD) δ ppm 0.93 (t, J=7.34 Hz, 3H) 0.97 (s, 9H) 1.08 (m, 2H) 1.24 (m,11H) 1.43 (m, 3H) 1.86 (m, 1H) 2.24 (m, 2H) 2.56 (m, 1H) 2.78 (q, J=7.09Hz, 2H) 2.92 (m, 1H) 4.01 (d, J=9.29 Hz, 1H) 4.22 (s, 1H) 4.29 (s, 1H)4.59 (d, J=6.85 Hz, 1H) 5.11 (d, J=10.76 Hz, 1H) 5.28 (d, J=17.36 Hz,1H) 5.49 (s, 1H) 5.74 (m, 1H) 7.66 (d, J=9.29 Hz, 1H) 8.18 (d, J=6.11Hz, 1H) 8.41 (m, 2H).

Example 205 Preparation of 5-propylthio-6-ethoxy isoquinoline P2*Derivatives

The following procedure is equally applicable to other5-alkylthio-6-alkoxy isoquinolines by changing the reagents shown here.To a solution of 1-chloro-6-fluoro isoquinoline (88 mg, 0.48 mmol) in2.0 mL THF under nitrogen at −78° C. was added LDA (1.5 Molar incyclohexane, 0.42 mL, 0.63 mmol) forming a dark brownish solution. Neatn-propyl disulfide (85 μL, excess) was introduced after it was stirredat −78° C. for 30 min. The reaction was allowed to warm to roomtemperature over a period of 30 min. It was quenched with a solution ofhalf saturated NH₄Cl, the organic residues were extracted into ethylacetate. The organic layers were combined and dried under vacuum to 50microns (Hg). The crude product was taken into 2 mL of THF, cooled to−78° C., added with excess potassium ethoxide (60 mg). The isoquinolineintermediate was finally purified by a silica gel column (type-H, Merck)eluted with ether-hexanes mixture, 32.2 mg (24%) of the pure compoundwas obtained. LC-MS showed 1-chloro-5-propylthio-6-ethoxyl isoquinolineat rt-min (MH⁺) [method C]: 3.77 (282). ¹H NMR (400 MHz, CHLOROFORM-D) δppm 0.94 (t, J=7.34 Hz, 3H) 1.46 (m, 2H) 1.55 (t, J=6.97 Hz, 3H) 2.83(t, J=7.21 Hz, 2H) 4.32 (q, J=6.85 Hz, 2H) 7.36 (d, J=9.29 Hz, 1H) 8.22(d, J=6.11 Hz, 1H) 8.32 (d, J=9.29 Hz, 1H) 8.35 (d, J=6.11 Hz, 1H).Following the general tripeptide alkylation procedure (Example 184),this 1-chloro-5-propylthio-6-ethoxy isoquinoline was alkylated with thetripeptide (compound 184) to give 40.7 mg (44.8%) of the desired productshown below.

Example 206 Preparation of Compound 206

BOCNH-P3(L-t-BuGly)-P2[(4R)-(6-ethoxy-5-propylthio-isoquinoline-1-oxo)-S-proline]-P1(1R,2SVinylAcca)-CONHSO₂ Cyclopropane. LC/MS rt-min (MH⁺): 2.93 (803) [methodB]. ¹H NMR (400 MHz, CD₃OD) δ ppm 0.93 (t, J=7.34 Hz, 3H) 1.01 (s, 9H)1.07 (m, 2H) 1.21 (m, 11H) 1.41 (m, 3H) 1.48 (t, J=6.85 Hz, 3H) 1.86(dd, J=8.07, 5.62 Hz, 1H) 2.25 (m, 2H) 2.60 (dd, J=13.69, 6.85 Hz, 1H)2.81 (q, J=6.97 Hz, 2H) 2.93 (m, 1H) 4.05 (m, 1H) 4.21 (s, 1H) 4.27 (q,J=7.09 Hz, 2H) 4.43 (d, J=11.74 Hz, 1H) 4.52 (m, 1H) 5.10 (d, J=10.76Hz, 1H) 5.28 (d, J=17.12 Hz, 1H) 5.74 (m, 1H) 5.82 (s, 1H) 7.30 (d,J=9.05 Hz, 1H) 7.92 (d, J=6.36 Hz, 1H) 7.97 (m, 1H) 8.22 (d, J=9.05 Hz,1H).

Example 207 Preparation of Compound 207

Likewise the same procedure was applied to the preparation ofBOCNH-P3(L-t-BuGly)-P2[(4R)-(6-methoxy-5-methylthio-isoquinolin-1-oxo)-S-proline]-P1(1R,2SVinylAcca)-CONHSO₂ Cyclopropane.

To the solution of 100 mg 1-Chloro-6-fluoro-isoquinoline (0.55 mmole) in2 ml dry THF at −78° C. was added LDA in THF (1.3 eq). Dark brownsolution was formed, then disulfide was added and the color of solutionchanged to greenish, then light brown. The reaction was quenched with 2mL of water and 2 mL of NH₄Cl, extracted with ethyl acetate, dried oversodium sulfate. The solvent was evaporated under vacuum and the resultedresidue was used as crude. LC/MS rt-min (MH⁺): 2.23 (228) [method B].The crude material was redissolved in 2 ml of dry THF at −78° C. and 1.3eq. of KOMe was added then the reaction mixture was allowed to warmed upto RT, stirred overnight. The reaction mixture was diluted with ethylacetate and washed with brine, dried over sodium sulfate. 104 mg wasobtained (79%). LC/MS rt-min (MH⁺): 2.04 (240) [method B]. Theintermediate, 1-chloro-5-methylthio-6-methoxy isoquinoline was subjectedto the tripeptide alkylation protocol described previously. Followingthe general procedure, 70.0 mg of yellow solid was obtained (42.7%).

LC/MS rt-min (MH⁺): 2.65 (760) [method B]. ¹H NMR (400 MHz,CHLOROFORM-D) δ ppm 0.94 (m, 11H) 1.17 (s, 9H) 1.26 (m, 2H) 1.39 (m, 1H)1.83 (dd, J=8.07, 5.62 Hz, 1H) 2.01 (m, 2H) 2.23 (s, 3H) 2.45 (m, 1H)2.79 (m, 1H) 3.94 (s, 3H) 3.97 (d, J=3.91 Hz, 1H) 4.15 (s, 1H) 4.25 (d,J=11.74 Hz, 1H) 4.36 (dd, J=9.66, 7.21 Hz, 1H) 4.99 (d, J=10.27 Hz, 1H)5.12 (d, J=16.87 Hz, 1H) 5.69 (m, 1H) 5.74 (s, 1H) 7.08 (d, J=9.05 Hz,1H) 7.83 (m, 2H) 8.06 (d, J=9.05 Hz, 1H).

Example 208 Preparation of1-chloro-6-methoxy-isoquinolin-5-yl-thiophen-2-yl-methanone

Following the same LDA deprotonation protocol (preparation of Example203) of 1-chloro-6-fluoro isoquinoline described previously, the initialanion was quenched with 2-thiophenecarboxaldehyde instead, to give1-chloro-6-fluoro isoquinolin-5-yl-thiophen-2-yl-methanol. The materialwas oxidized to the1-chloro-6-fluoro-isoquinolin-5-yl-thiophen-2-yl-methanone using MnO₂ inbenzene in 49.6% overall yield after chromatographic purification. LC/MSrt-min (MH⁺) [method C]: 2.98 (292). ¹H NMR (400 MHz, CHLOROFORM-D) δppm 7.12 (dd, J=4.89, 3.91 Hz, 1H) 7.40 (m, 1H) 7.53 (m, 1H) 7.56 (dd,J=5.87, 0.73 Hz, 1H) 7.82 (dd, J=5.01, 1.10 Hz, 1H) 8.27 (d, J=5.87 Hz,1H) 8.54 (ddd, J=9.29, 5.38, 0.73 Hz, 1H). Ipso nucleophilic aromaticdisplacement of the fluorine atom was accomplished in a solution ofexcess of potassium methoxide to give, mainly1-chloro-6-methoxy-isoquinolin-5-yl-thiophen-2-yl-methanone along with25-33% of 1,6-dimethoxy-isoquinolin-5-yl-thiophen-2-yl-methanone. Thecrude material (77 mg) was used in the alkylation step with thetripeptide without further purification.

Example 209 Preparation of Compound 209

Following the general procedure of tripeptide alkylation (Example 184),35.3 mg ofBOCNH-P3(L-t-BuGly)-P2[(4R)-6-methoxy-5-(thiophene-2-carbonyl)-isoquinoline-1-oxo)-S-proline]-P1(1R,2SVinylAcca)-CONHSO₂ Cyclopropane was obtained as pale solid (26.5%).LC/MS rt-min (MH⁺): 2.54 (825) [method B]. ¹H NMR (400 MHz, CD₃OD) δ ppm1.02 (s, 9H) 1.06 (m, 2H) 1.22 (m, 2H) 1.26 (s, 9H) 1.43 (m, 1H) 1.87(dd, J=7.95, 5.50 Hz, 1H) 2.28 (m, 2H) 2.62 (dd, J=13.82, 6.97 Hz, 1H)2.93 (m, 1H) 3.90 (s, 3H) 4.07 (dd, J=11.62, 3.06 Hz, 1H) 4.23 (s, 1H)4.43 (m, 1H) 4.55 (dd, J=9.78, 7.34 Hz, 1H) 5.09 (m, 1H) 5.29 (d,J=17.12 Hz, 1H) 5.74 (m, 1H) 5.86 (s, 1H) 6.93 (d, J=6.11 Hz, 1H) 7.11(m, 1H) 7.32 (dd, J=3.91, 0.98 Hz, 1H) 7.46 (d, J=9.29 Hz, 1H) 7.86 (t,J=6.72 Hz, 1H) 7.91 (dd, J=4.89, 1.22 Hz, 1H) 8.39 (d, J=9.29 Hz, 1H).

Preparation of P2* by way of Cinnamic Acid Derivatives. The GeneralProcedure Depicted Below has been Described Extensively Elsewhere.

Example 210 Preparation of Compound 210

10.0 g of meta-tolyl-acrylic acid (61.7 mmole) was suspended in 50 ml ofbenzene, 12.6 mL of DPPA (0.95 eq) was added followed by 10.3 ml oftriethylamine (1.2 eq). The resulted solution was stirred at roomtemperature for 1 hr. The volatile was removed under vacuum and themeta-tolyl-acryloyl azide was purified by flash chromatograph to yield11.5 g of pure compound (quantitative). This material, in 100 mL ofdiphenylmethane, was introduced dropwise into 100 ml of diphenylmethanepreviously heated up to 200° C. over a period of an hr. The resultedsolution was kept at this temperature for another 4 hour then cooleddown to room temp. White precipitate was formed, it was filtered off.The solid was washed with hexanes three times and dried. The filtratewas diluted with 200 ml of hexanes, the solution was left standingovernight to allow for separation of the second crop. The materials werecombined to give 4.2 g of 6-methyl-isoquinolin-1-ol (50%). LC/MS rt-min(MH⁺): 1.31 (160) [method B]. ¹H NMR (400 MHz, CD₃OD) δ ppm 2.49 (s, 3H)6.61 (d, J=7.32 Hz, 1H) 7.13 (d, J=7.02 Hz, 1H) 7.36 (d, J=8.24 Hz, 1H)7.45 (s, 1H) 8.18 (d, J=8.24 Hz, 1H). The material was suspended in 15ml of POCl₃ and brought to reflux for 3 hours. After removal of thePOCl₃ in vacuo, the residue was partitioned between EtOAc (1 L), andcold aqueous NaOH (generated from 1.0N 200 mL NaOH and 20 mL 10.0 NNaOH) and stirred for 15 min. The organic layer was washed with water(2×200 mL), brine (200 mL), dried (MgSO₄), and concentrated in vacuo tosupply 1-chloro-6-methyl-isoquinoline (67.4%). LC/MS rt-min (MH⁺): 1.92(178) [method B]. ¹H NMR (400 MHz, CHLOROFORM-D) δ ppm 2.53 (s, 3H) 7.47(d, J=6.11 Hz, 2H) 7.56 (s, 1H) 8.18 (m, 2H). The final alkylation of1-chloro-6-methyl-isoquinoline with the tripeptide was carried out usingthe protocol described previously (Example 184).

BOCNH-P3(L-tert-BuGly)-P2[(4R)-(6-methylisoquinolin-1-oxo)-S-proline]-P1(1R,2S Vinyl Acca)-CONHSO₂-Cyclopropane:the material was obtained as a white foam in 18% yield. LC/MS R_(t)-min(MNa⁺) [method B]: 2.64 (720). ¹H NMR (400 MHz, CD₃OD) δ ppm 1.05 (m,13H) 1.23 (m, 9H) 1.42 (m, 1H) 1.86 (dd, J=7.95, 5.50 Hz, 1H) 2.25 (m,2H) 2.49 (s, 3H) 2.61 (dd, J=13.82, 6.48 Hz, 1H) 2.93 (m, 1H) 4.05 (dd,J=11.86, 3.30 Hz, 1H) 4.23 (s, 1H) 4.43 (d, J=11.49 Hz, 1H) 4.52 (m, 1H)5.10 (d, J=11.49 Hz, 1H) 5.28 (d, J=17.12 Hz, 1H) 5.74 (m, 1H) 5.83 (s,1H) 7.24 (d, J=5.87 Hz, 1H) 7.35 (d, J=8.07 Hz, 1H) 7.58 (s, 1H) 7.89(d, J=5.87 Hz, 1H) 8.07 (d, J=8.56 Hz, 1H).

Example 211 Preparation of Compound 211

Following the general procedure described previously,BOCNH-P3(L-t-BuGly)-P2[(4R)-(1,3-Dioxa-7-aza-cyclopenta[a]naphthalen-6-ol)-S-proline]-P(1R,2S VinylAcca)-CONHSO₂ Cyclopropane, 51.0 mg was obtained as a palesolid (64.9%). LC/MS rt-min (MH⁺): 2.57 (728) [method B]. ¹H NMR (400MHz, CD₃OD) δ ppm 0.99 (s, 9H) 1.07 (m, 2H) 1.18 (m, 11H) 1.42 (m, 1H)1.86 (dd, J=8.07, 5.62 Hz, 1H) 2.23 (m, 2H) 2.59 (dd, J=13.69, 6.85 Hz,1H) 2.93 (m, 1H) 4.04 (dd, J=11.74, 2.20 Hz, 1H) 4.22 (s, 1H) 4.43 (d,J=11.74 Hz, 1H) 4.51 (m, 1H) 5.10 (d, J=10.27 Hz, 1H) 5.28 (d, J=17.12Hz, 1H) 5.73 (m, 1H) 5.82 (s, 1H) 6.18 (s, 2H) 7.13 (d, J=8.56 Hz, 1H)7.19 (d, J=6.11 Hz, 1H) 7.81 (d, J=8.56 Hz, 1H) 7.85 (d, J=6.11 Hz, 1H).

Preparation of 5,6,7-Trisubstituted Isoquinoline P2* Derivatives

The 5,6-methylenedioxy-1-chloroisoquinoline prepared above wasdeprotonated directly in the presence of strong base such as LDA toprovide the corresponding 7-anion without interfering with the 1-chlorofunctionality. The 7-anion was quenched with electrophiles such as NFSI(N-fluorobenzenesulfonimide) and dimethylsulfide to produce thecorresponding 7-substituted isoquinoline ring system.

Example 212 Preparation of Compound 212

Step 1:

Preparation of 5,6-methylenedioxy-7-fluoro-1-chloro isoquinoline. To asolution of 5,6-methylenedioxy-1-chloro isoquinoline (126 mg, 0.61 mmol)in 4 mL of THF under nitrogen at −78° C. was added LDA solution incyclohexane (1.5 Molar, 0.65 mL, 0.98 mmol). The light brownish solutionwas stirred for 15 min before it was treated withN-fluorobenzenesulfonimide (NFSI, 0.3 g, 1.5 equivalents). TLC analysisshowed the formation of a new spot, in addition of the unchangedstarting material. Aqueous work up followed by extractions with ethylacetate furnished an oily crude product which was purified bypreparative HPLC to give 52 mg (38%). LC/MS rt-min (MH⁺) [method C]:3.09 (226). ¹H NMR (400 MHz, CHLOROFORM-D) δ ppm 6.33 (s, 2H) 7.52 (d,J=5.87 Hz, 1H) 7.71 (d, J=10.51 Hz, 1H) 8.16 (d, J=5.87 Hz, 1H). Step 2:The alkylation of 5,6-methylenedioxy-7-fluoro-1-chloro isoquinoline withthe tripeptide was carried out as described previously (Example 184) toprovide the major product as a result of fluorine displacement.

BOCNH-P3(L-t-BuGly)-P2[(4R)-(6-Chloro-1,3-dioxa-7-aza-cyclopenta[a]naphthalen-4-oxo)-S-proline]-P1(1R,2SVinylAcca)-CONHSO₂ Cyclopropane, Shown below

Following the general procedure, 24.3 mg of yellow solid was obtained(24.3%). LC/MS rt-min (MH⁺): 2.54 (763) [method B]. ¹H NMR (400 MHz,CD₃OD) δ ppm 1.00 (s, 9H) 1.06 (m, 2H) 1.20 (m, 2H) 1.29 (s, 9H) 1.42(dd, J=9.41, 5.26 Hz, 1H) 1.86 (dd, J=8.07, 5.38 Hz, 1H) 2.24 (m, 2H)2.54 (dd, J=13.57, 6.48 Hz, 1H) 2.92 (m, 1H) 4.04 (dd, J=12.10, 2.81 Hz,1H) 4.20 (d, J=7.34 Hz, 1H) 4.33 (d, J=12.23 Hz, 1H) 4.47 (dd, J=10.52,6.85 Hz, 1H) 5.10 (dd, J=10.39, 1.59 Hz, 1H) 5.28 (dd, J=17.12, 1.22 Hz,1H) 5.46 (d, J=5.87 Hz, 1H) 5.74 (m, 1H) 6.29 (m, 2H) 7.40 (s, 1H) 7.56(m, 1H) 8.01 (d, J=5.62 Hz, 1H).

Example 213 Preparation of Compound 213

To a solution of 5,6-methylenedioxy-1-chloro isoquinoline (84 mg, 0.41mmol) in 4 mL of THF under nitrogen at −78° C. was added LDA solution incyclohexane (1.5 Molar, 0.60 mL, 0.9 mmol). The light brownish solutionwas stirred for 15 min at −78° C. before it was treated with methyldisulfide (50 μL of neat reagent, 1.4 equivalents). TLC analysis showedthe formation of a new spot, in addition of the unchanged startingmaterial. Aqueous work up followed by extractions with ethyl acetatefurnished an oily crude product which was purified by preparative HPLCto give 51 mg (49%). LC/MS rt-min (MH⁺) [method C]: 3.39 (254). ¹H NMR(400 MHz, CHLOROFORM-D) δ ppm 2.64 (s, 3H) 6.29 (s, 2H) 7.49 (d, J=4.89Hz, 1H) 7.71 (s, 1H) 8.11 (d, J=5.87 Hz, 1H). The alkylation of5,6-methylenedioxy-7-methylthio-1-chloro isoquinoline with thetripeptide was carried out as described previously (Example 184) toprovide the desired product shown below:

BOCNH-P3(L-t-BuGly)-P2[(4R)-(4-methylthio-1,3-dioxa-7-aza-cyclopenta[a]naphthalen-6-yloxy)-S-proline]-P1(1R,2SVinylAcca)-CONHSO₂ Cyclopropane

Following the general procedure, 59.6 mg of yellow solid was obtained(42.2%). LC/MS rt-min (MH⁺): 2.70 (774) [method B]. ¹H NMR (400 MHz,CHLOROFORM-D) δ ppm 1.02 (m, 11H) 1.16 (s, 9H) 1.32 (s, 2H) 1.45 (m, 1H)1.94 (m, 1H) 2.12 (d, J=8.56 Hz, 1H) 2.56 (s, 3H) 2.62 (m, 2H) 2.90 (d,J=4.40 Hz, 1H) 4.15 (d, J=7.83 Hz, 2H) 4.48 (d, J=12.47 Hz, 1H) 4.62 (t,J=7.83 Hz, 1H) 5.13 (d, J=10.52 Hz, 1H) 5.26 (d, J=17.12 Hz, 1H) 5.74(d, J=16.38 Hz, 1H) 5.95 (s, 1H) 6.28 (s, 2H) 7.41 (s, 1H) 7.59 (s, 1H)7.85 (d, J=6.11 Hz, 1H).

Preparation of 3,4-disubstituted Isoquinoline P2* Derivatives Example215 Preparation of Compound 215

BOCNH-P3(L-t-BuGly)-P2[(1R)-(2,3-dihydro-1H-4-aza-cyclopenta[a]naphthalen-5-yloxy)-S-proline]-P1(1R,2SVinylAcca)-CONHSO₂ Cyclopropane, shown below was prepared as depicted inthe following scheme:

Notes:

The synthesis of the novel 1-fluoro P2* was made successful by thetechnologies cited below:

-   (1) Rigby, James H.; Holsworth, Daniel D.; James, Kelly. Vinyl    Isocyanates In Synthesis. [4+2] Cycloaddition Reactions With Benzyne    Addends. Journal Of Organic Chemistry (1989), 54(17), 4019-20-   (2) Uchibori, Y.; Umeno, M.; Yoshiokai, H.; Heterocycles, 1992, 34    (8), 1507-1510

Example 214 Preparation of Compounds 214,5-chloro-2,3-dihydro-1H-4-aza-cyclopenta[a]naphthalene, and Compound 215of Example 215

2,3-Dihydro-1H-4-aza-cyclopenta[a]naphthalen-5-ol was prepared inaccordance to the method of Rigby described in (reference 1) citedabove. Using POCl₃ as described elsewhere, Compound 214, was synthesizedin 59.8% (430 mg). LC/MS rt-min (MH⁺): 2.29 (204) [method B]. ¹H NMR(400 MHz, CHLOROFORM-D) ppm 2.28 (m, 2H) 3.19 (q, J=7.74 Hz, 4H) 7.58(m, 1H) 7.71 (m, 2H) 8.32 (d, J=8.56 Hz, 1H). The chloride issufficiently reactive to be alkylated with the tripeptide according tothe procedure of Example 184, to give the desired product Compound 215.However the overall yield could be doubled if the chloride was exchangedinto the fluoride by the method of Uchibori described in (reference 2).Thus 17.0 mg of Compound 215, isolated as a pale yellow solid (23.6%).LC/MS rt-min (MH⁺): 2.80 (724) [method B]. ¹H NMR (500 MHz, CD₃OD) δ ppm1.03 (s, 9H) 1.09 (m, 2H) 1.24 (m, 11H) 1.44 (dd, J=8.24, 5.49 Hz, 1H)1.88 (dd, J=7.93, 5.49 Hz, 1H) 2.25 (m, 4H) 2.63 (dd, J=13.73, 7.02 Hz,1H) 2.94 (m, 1H) 3.05 (m, 2H) 3.10 (m, 2H) 4.08 (dd, J=11.60, 2.75 Hz,1H) 4.24 (d, J=20.45 Hz, 1H) 4.45 (d, J=11.90 Hz, 1H) 4.54 (dd, J=9.46,7.63 Hz, 1H) 5.11 (m, 1H) 5.30 (d, J=17.09 Hz, 1H) 5.75 (m, 1H) 5.87 (s,1H) 7.44 (t, J=7.02 Hz, 1H) 7.69 (m, 2H) 8.18 (d, J=8.24 Hz, 1H).

Preparation of 3,4-dihydrofuranyl and Furanyl Isoquinoline P2*Components, Examples 217 and 218

Example 217 Preparation of Compound 217,5-chloro-2,3-dihydro-1-oxa-4-aza-cyclopenta[a]naphthalene and Compound218, 5-chloro-1-oxa-4-aza-cyclopenta[a]naphthalene

This synthesis made use of the technologies described, in part, in thefollowing references:

-   (1) Hojo, Masaru; Masuda, Ryoichi; Sakaguchi, Syuhei; Takagawa,    Makoto, Synthesis (1986), (12), 1016-17-   (2) Rigby, James H.; Holsworth, Daniel D.; James, Kelly. Vinyl    Isocyanates In Synthesis. [4+2] Cycloaddition Reactions With Benzyne    Addends. Journal Of Organic Chemistry (1989), 54(17), 4019-20-   (3) Uchibori, Y.; Umeno, M.; Yoshiokai, H.; Heterocycles, 1992, 34    (8), 1507-1510

Both 2,3-dihydro-1-oxa-4-aza-cyclopenta[a]naphthalen-5-ol and1-oxa-4-aza cyclopenta[a]naphthalen-5-ol were produced together when theprocedures (references 1 and 2) cited above were followed. Conversion ofthe pair into their chloro derivatives was accomplished by POCl₃ asusual: The crude hydroxy products (about 2 g, pale yellow oil) wastreated with 15 mL of POCl₃ and the mixture was brought to reflux for 3hours. After removal of the POCl₃ in vacuo, the residue was stirred withEtOAc (1 L), and cold aqueous NaOH (220 mL, 1.0 N) for 15 min. Theorganic layer was separated, washed with water (2×200 mL), brine (200mL), and dried over MgSO₄, and concentrated in vacuo to supply 300 mg ofExample 217, 5-chloro-2,3-dihydro-1-oxa-4-aza-cyclopenta[a]naphthalene(13.2%) and 100 mg of Example 218, 5-chloro-1-oxa-4-azacyclopenta[a]naphthalene (4.4%) as light brown solids after silica gelchromatographic separation. Compound 217: LC/MS rt-min (MH⁺): 2.05 (206)[method B]. ¹H NMR (400 MHz, CHLOROFORM-D) δ ppm 3.46 (t, J=9.05 Hz, 2H)4.82 (t, J=9.17 Hz, 2H) 7.58 (m, 1H) 7.66 (m, 1H) 7.85 (d, J=8.31 Hz,1H) 8.21 (d, J=8.56 Hz, 1H). Compound 218: LC/MS rt-min (MH⁺): 2.16(204) [method B]. ¹H NMR (400 MHz, CHLOROFORM-D) ppm 7.15 (d, J=2.20 Hz,1H) 7.70 (m, 1H) 7.89 (m, 2H) 8.27 (d, J=8.31 Hz, 1H) 8.44 (d, J=8.80Hz, 1H).

Preparation of5-fluoro-2,3-dihydro-1-oxa-4-aza-cyclopenta[a]naphthalene, and Final P2*Coupling Products

The chloride/fluoride exchange was achieved by the method (reference 3)cited above. Thus 90 mg of5-chloro-2,3-dihydro-1-oxa-4-aza-cyclopenta[a]naphthalene (Example 217)was suspended in 1.5 mL of Bu₄PHF₂ and was irradiated under microwave(Smith Reactor) to about 120° C. for 2 hours. After aqueous work up andcolumn purification, 22 mg of fluoride product was obtained (26.9%).LC/MS rt-min (MH⁺): 1.91 (190) [method B]. The furan derivative (Example218), 5-chloro-1-oxa-4-aza-cyclopenta[a]naphthalene was sufficientlyreactive to be alkylated with the tripeptide directly without fluorideactivation.

Example 219 Preparation of Compound 219

BOCNH-P3(L-t-BuGly)-P2[(4R)-(2,3-dihydro-1-oxa-4-aza-cyclopenta[a]naphthalen-5-yloxy)-S-proline]-P1(1R,2SVinylAcca)-CONHSO₂ Cyclopropane. Following the general alkylationprocedure, 22.0 mg of yellow solid was obtained (26.3%). LC/MS rt-min(MH⁺): 2.65 (726) [method B]. ¹H NMR (400 MHz, CD₃OD) δ ppm 0.99 (s, 9H)1.07 (m, 2H) 1.20 (m, 11H) 1.40 (m, 1H) 1.86 (dd, J=8.07, 5.62 Hz, 1H)2.21 (dd, J=17.48, 8.93 Hz, 2H) 2.60 (dd, J=13.45, 6.85 Hz, 1H) 2.93 (m,1H) 3.34 (m, 2H) 4.04 (dd, J=11.74, 3.18 Hz, 1H) 4.24 (s, 1H) 4.41 (d,J=11.49 Hz, 1H) 4.51 (m, 1H) 4.74 (t, J=9.05 Hz, 2H) 5.11 (d, J=10.27Hz, 1H) 5.28 (d, J=17.36 Hz, 1H) 5.73 (m, 1H) 5.78 (s, 1H) 7.43 (m, 1H)7.65 (t, J=7.46 Hz, 1H) 7.74 (d, J=8.31 Hz, 1H) 8.12 (d, J=8.56 Hz, 1H).

Example 220 Preparation of Compound 220

BOCNH-P3(L-t-BuGly)-P2[(4R)-(1-oxa-4-aza-cyclopenta[a]naphthalen-5-yloxy)-S-proline]-P1(1R,2SVinylAcca)-CONHSO₂ Cyclopropane. Following the general alkylationprocedure, 13.0 mg of yellow solid was obtained (20%). LC/MS rt-min(MH⁺): 2.70 (724) [method B]. ¹H NMR (500 MHz, CD₃OD) δ ppm 1.01 (S, 9H)1.09 (m, 2H) 1.22 (s, 9H) 1.27 (m, 2H) 1.46 (m, 1H) 1.89 (dd, J=7.78,5.65 Hz, 1H) 2.24 (d, J=8.55 Hz, 1H) 2.33 (t, J=9.92 Hz, 1H) 2.68 (dd,J=13.73, 7.02 Hz, 1H) 2.95 (m, 1H) 4.14 (m, 1H) 4.26 (s, 1H) 4.50 (d,J=11.90 Hz, 1H) 4.57 (d, J=17.09 Hz, 1H) 5.12 (d, J=10.07 Hz, 1H) 5.30(d, J=17.40 Hz, 1H) 5.75 (m, 1H) 5.93 (s, 1H) 6.97 (d, J=2.14 Hz, 1H)7.51 (t, J=7.32 Hz, 1H) 7.81 (t, J=7.48 Hz, 1H) 7.92 (s, 1H) 8.13 (d,J=7.94 Hz, 1H) 8.28 (d, J=8.24 Hz, 1H).

Preparation of 3-halo and 3-heteroaryl 4-alkoxy and 4-hydroxyisoquinoline P2* Derivatives

4-Methoxy isoquinoline (Example 222a) was prepared from 4-bromoisoquinoline by a novel and convenient procedure using ordinarylaboratory equipments and reagents. A regioselective NBS brominationgave 3-bromo-4-methoxy isoquinoline (Example 222b) in good yields. MCPBAoxidation proceeded uneventfully to furnish the corresponding N-oxide(Example 222c), which was isomerized into 1-chloro-3-bromo-4-methoxyisoquinoline (Example 222d) using the usual POCl₃ procedure. The4-methoxy isoquinoline was alkylated with the tripeptide to give thecorresponding 3-bromo-4-methoxy P2* derivative suitable for Stille andSuzuki coupling. Alternatively the 4-methoxy isoquinoline wasde-methylated in BBr₃ to give the 4-hydroxy-3-bromo-1-chloroisoquinoline (Example 222e). The 4-hydroxy group was re-protected withSEM-chloride to give the 4-SEM protected intermediate Example 222. The4-hydroxy compound was re-generated once the coupling was achieved byeither an acid induced, or a fluoride induced deprotection protocol.

Example 222d Preparation of 1-chloro-3-bromo-4-methoxy isoquinoline

Step 1:

A solution of 4-bromo isoquinoline (15 g, 73 mmol, commercial material)in 200 mL dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone (DMPU, Aldrich)was added solid potassium methoxide (5.6 gm, 80 mmol). The reactionvessel was immersed in an oil bath at 105° C. for 20 min. The color ofmixture changed rapidly from its initial very pale to dark greenishbrown immediately after warming. The reaction vessel was removed fromthe oil bath and was diluted with water, the organic residues werepartitioned into ether by multiple extraction with portions of ether.TLC analysis showed two new, spots (1:1 v/v mixture of hexanes and ethylacetate as eluent) of roughly equal size. These were separated onsilica-gel (Merck, type-H) column eluted with straight hexanes, followedby gradual addition of ether into the mobile phase. The desired product,4-methoxy isoquinoline (4.1 gm, 35.3%) was isolated after evaporation ofsolvents. The other product was also isolated as the reductionby-product isoquinoline. The identity of the by-product was confirmed byNMR comparison with authentic material. Example 222a: LC/MS R_(t)-min(MH⁺) [method C]: 1.16 (160). ¹H NMR (400 MHz, CHLOROFORM-D) δ 4.07 (s,3H) 7.61 (m, 1H) 7.69 (m, 1H) 7.93 (d, J=8.07 Hz, 1H) 8.08 (s, 1H) 8.19(d, J=8.56 Hz, 1H) 8.89 (s, 1H). [Note: this compound was previouslyprepared in Zoltewicz, John A.; Oestreich, Terence M.; Sale, Alan A,Journal of the American Chemical Society (1975), 97(20), 5889-96 in a“Monel Bomb”, and later by a “focused microwave” initiated procedure inChemg, Yie-Jia, Tetrahedron (2002), 58(6), 1125-1129. The presentprocedure required neither special high pressure apparatus norpreparative scale microwave equipment].

Step 2:

The material (Example 222a) was subjected to NBS bromination, thus4-methoxy isoquinoline (Example 222a, 2.1 gm, 13.2 mmol) in1,2-dichloroethane (DCE, 150 mL) was treated with N-bromosuccinimide(NBS, 1.5 gm, 8.4 mmol, 0.6×) at 70° C. for an hr followed by additionof second portion of 1.5 gm NBS. The dark brownish mixture was stirredfor another hr before the addition of third portion of 1.0 gm NBS. Thebromination was monitored by LC-MS until there was no starting materialleft. The crude mixture was evaporated to dryness and the desiredproduct was filtered over a short bed of silica-gel (Type-H, Merck, 3 cmdiameter by 1.5 cm height) eluted with straight hexanes first followedby gradually increasing the amount of ether. The desired product,(Example 222b), was isolated as an oily material (1.7 gm, 54%). LC/MSR_(t)-min (MH⁺) [method C]: 2.65 (238). ¹H NMR (400 MHz, CHLOROFORM-D) δppm 4.04 (s, 3H) 7.64 (t, J=7.58 Hz, 1H) 7.76 (t, J=7.09 Hz, 1H) 7.99(d, J=8.31 Hz, 1H) 8.11 (d, J=8.31 Hz, 1H) 8.85 (s, 1H).[3-Bromo-4-methoxy isoquinoline was previously prepared by a differentprocedure: Finkentey, Christel; Langhals, Elke; Langhals, Heinz.Chemische Berichte (1983), 116(6), 2394-7. NMR of the product wasidentical to that reported].

Step 3:

The product from NBS bromination was subjected to MCPBA oxidation inmethylene chloride at room temperature. Thus MCPBA (1.80 gm, 77% pure,8.0 mmol) was added into a solution of 3-bromo-4-methoxy isoquinoline(Example 222b, 1.65 gm, 6.9 mmol) in 35 mL of CH₂Cl₂. The solution wasstirred for 4 hrs forming a white suspension. Sodium bicarbonatesolution (5%, freshly prepared, 20 mL) was added into the mixture,organic residues were extracted into CH₂Cl₂ (10×25 mL). Multipleextraction in organic solvent was necessary to recover the somewhatwater solution N-oxide product. The crude material obtained afterevaporation of solvents was further purified by a filtration oversilica-gel to give 1.36 gm (5.4 mmol, 78%) of the N-oxide (Example 222c)as a ceraceous solid.

LC/MS R_(t)-min (MH⁺) [method C]: 1.79 (254). ¹H NMR (400 MHz,CHLOROFORM-D) δ ppm 4.07 (s, 3H) 7.63 (m, 2H) 7.72 (m, 1H) 8.00 (m, 1H)8.86 (s, 1H).

Step 4:

The final N-oxide rearrangement was done as usual in POCl₃ usingprocedure described elsewhere. Yield of Example 222d was essentiallyquantitative. LC/MS R_(t)-min (MH⁺) [method D]: 2.69 (272). ¹H NMR asHCl salt, (400 MHz, CHLOROFORM-D) δ ppm 4.07 (s, 3H) 7.81 (m, 1H) 7.92(m, 1H) 8.17 (d, J=8.31 Hz, 1H) 8.34 (d, J=8.31 Hz, 1H). ¹H NMR as freebase, (400 MHz, CHLOROFORM-D) δ ppm 4.03 (s, 3H) 7.72 (m, 1H) 7.81 (m,1H) 8.12 (d, J=8.56 Hz, 1H) 8.28 (d, J=8.56 Hz, 1H).

Example 223 Preparation of Compound 223

The free base (Example 222d) obtained in the previous step was alkylatedwith the tripeptide fragment using the alkylation protocol (Example 184)described elsewhere to give 79% of the desired product as a paper-whitesolid. LC/MS R_(t)-min (MNa⁺) [method C]: 3.91 (814). ¹H NMR (400 MHz,CD₃OD) δ ppm 1.02 (s, 9H) 1.06 (dd, J=8.07, 1.47 Hz, 2H) 1.22 (m, 11H)1.42 (dd, J=9.78, 5.14 Hz, 1H) 1.86 (dd, J=8.07, 5.38 Hz, 1H) 2.22 (dd,J=18.10, 9.29 Hz, 1H) 2.28 (m, 1H) 2.61 (dd, J=13.57, 6.97 Hz, 1H) 2.93(m, 1H) 3.92 (s, 3H) 4.06 (dd, J=11.86, 2.81 Hz, 1H) 4.22 (s, 1H) 4.43(d, J=11.49 Hz, 1H) 4.51 (m, 1H) 5.10 (d, J=10.52 Hz, 1H) 5.28 (d,J=17.12 Hz, 1H) 5.74 (m, 1H) 5.81 (s, 1H) 7.56 (t, J=7.58 Hz, 1H) 7.78(t, J=7.58 Hz, 1H) 8.00 (d, J=8.31 Hz, 1H) 8.16 (d, J=8.56 Hz, 1H).

Example 224 and 225 Preparation of Compounds 224 and Compound 225

The 4-methoxy group in 1-chloro-3-bromo-4-methoxy isoquinoline (Example222d) described previously was converted into α-trimethylsilyl ethoxymethyl (SEM) moiety by the following procedure.1-Chloro-3-bromo-4-methoxy isoquinoline (Example 222d) was demethylatedusing BBr₃ (final adjusted reaction concentration was 0.2-0.3 MolarBBr₃) at room temperature for 12 hrs. The high BBr₃ concentration forsuch de-methylation was found to be necessary and efficient. The crudereaction mixture was diluted with 50 volumes of anhydrous methanol priorto evaporation to dryness. The demethylation was essentiallyquantitative. Example 222e: LC/MS R_(t)-min (MH⁺) [method D]: 2.32(258). ¹H NMR of free HCl salt (400 MHz, CHLOROFORM-D) δ ppm 5.83 (br.s, 1H) 7.73 (t, J=7.70 Hz, 1H) 7.79 (t, J=7.58 Hz, 1H) 8.22 (m, 2H). The4-hydroxy-3-bromo-1-chloro isoquinoline (Example 222e) was re-protectedwith 2-(trimethylsilyl)ethoxy methyl chloride (SEM-Cl). The crude freebase from the previous preparation was dried to 40 microns (Hg) at roomtemperature prior to re-protection with SEM-chloride. To a solution ofthe 4-hydroxy compound (Example 222e, 1.33 gm, 5.2 mmol) in methylenechloride (50 ml) at 0° C. was added sequentially diisopropylethyl amine(2 mL, 11.5 mmol) and SEM-chloride (1.8 mL, 10 mmol). The mixture wasstirred for min before it was washed with a freshly prepared NaHCO₃solution (5%, 100 mL). The organic residues were extracted into severalportions of methylene chloride, the combined organic layers wereback-washed with 20 mL deionized water before it was concentrated invacuo. The SEM protection was essentially quantitative. Example 222LC/MS R_(t)-min (MH⁺) [method D]: 3.40 (410). ¹H NMR (400 MHz,CHLOROFORM-D) δ ppm 0.03 (s, 9H) 0.99 (m, 2H) 3.98 (m, 2H) 5.33 (s, 2H)7.72 (m, 1H) 7.80 (m, 1H) 8.17 (d, J=8.56 Hz, 1H) 8.27 (d, J=8.07 Hz,1H). Alkylation of 4-SEM protected isoquinoline with tripeptide:Compound 224 and Compound 225 were generated from the same tripeptidealkylation reaction. The 4-hydroxy compound (Compound 224) was producedmost probably as a result of the TFA present during the preparative HPLCpurification.

Example 224 (15.4%): LC/MS R_(t)-min (MNa⁺) [method D]: 2.87 (800). ¹HNMR (400 MHz, CD₃OD) δ ppm 1.02 (s, 9H) 1.06 (d, J=8.31 Hz, 2H) 1.25 (s,9H) 1.42 (s, 2H) 1.86 (m, 1H) 2.23 (m, 2H) 2.60 (dd, J=13.21, 7.34 Hz,1H) 2.93 (m, 1H) 4.06 (d, J=11.00 Hz, 1H) 4.24 (m, 2H) 4.38 (d, J=11.98Hz, 1H) 4.49 (dd, J=9.78, 7.09 Hz, 1H) 5.10 (d, J=10.03 Hz, 1H) 5.28 (d,J=17.36 Hz, 1H) 5.72 (m, 1H) 5.76 (s, 1H) 7.52 (t, J=7.46 Hz, 1H) 7.71(t, J=7.09 Hz, 1H) 8.09 (d, J=4.40 Hz, 1H) 8.11 (d, J=4.16 Hz, 1H).

Example 225 (8.0%): LC/MS R_(t)-min ([M-BOC]⁺) [method D]: 3.46 (808).¹H NMR (400 MHz, CD₃OD) δ ppm 0.01 (s, 9H) 0.96 (m, 2H) 1.02 (s, 11H)1.06 (d, J=6.60 Hz, 2H) 1.24 (s, 9H) 1.42 (m, 1H) 1.86 (dd, J=7.83, 5.38Hz, 1H) 2.25 (m, 2H) 2.62 (dd, J=13.69, 7.34 Hz, 1H) 2.93 (m, 1H) 3.97(m, 2H) 4.07 (dd, J=10.88, 3.55 Hz, 1H) 4.23 (s, 1H) 4.43 (d, J=11.25Hz, 1H) 4.50 (m, 1H) 5.10 (d, J=10.76 Hz, 1H) 5.25 (m, 3H) 5.74 (m, 1H)5.82 (s, 1H) 7.57 (m, 1H) 7.77 (t, J=7.83 Hz, 1H) 8.06 (d, J=8.56 Hz,1H) 8.16 (d, J=8.31 Hz, 1H).

Preparation of 4H-[1,3]dioxino[5,4-c]isoquinolin P2* derivatives

Reaction Conditions: (1) MeOK in DMPU; (2) MCPBA in CH₂Cl₂; (3) POCl₃ inDCE; (4) BBr₃ in CH₂Cl₂; (5) HCHO solution in 40% H₂SO₄ by procedure ofSynthesis of 1,3-oxazino[5,6-c]isoquinolines and related compounds.Miyoko Toyama and Hirotaka Otomasu, Chem. Pharm. Bull. 33(12),5543-5546, 1985; (6) Fluororination procedure by Uchibori, Y.; Umeno,M.; Yoshiokai, H.; Heterocycles, 1992, 34 (8), 1507-1510

Example 227 Preparation of Compound 227

6-Chloro-1,3-Oxazino[5,6-c]isoquinoline was prepared by the procedure ofMiyoko Toyama and Hirotaka Otomasu starting from 1-chloro-4-hydroxyisoquinoline. The starting material: 1-chloro-4-hydroxy isoquinoline(Example 226c) was prepared by the synthetic sequence shown above. MCPBAoxidation of 4-methoxy isoquinoline (Example 222a) was carried as usualto give 79.1% of the corresponding N-oxide (Example 226a). The materialwas converted into the 1-chloro derivative immediately afterward inPOCl₃ to give the chloride (Example 226b) in essentially quantitativeyield. The crude 1-chloro-4-methoxy isoquinoline was de-methylated inBBr₃ at room temperature to give the corresponding 1-chloro-4-hydroxyisoquinoline (Example 226c) after treating the crude BBr₃ mixture withanhydrous methanol at room temperature, followed by evaporation to getrid of excess of borate residues. The reaction of Miyoko Toyama andHirotaka Otomasu gave 266 mg of 6-chloro-1,3-oxazino[5,6-c]isoquinoline(Example 226d, 62.3%) overall yield from 300 mg of 4-methoxyisoquinoline in 4 steps. LC/MS R_(t)-min ([M−HCHO]H⁺) [method D]: 2.45(192). ¹H NMR (400 MHz, CHLOROFORM-D) δ ppm 5.02 (s, 2H) 5.41 (s, 2H)7.68 (m, 1H) 7.77 (ddd, J=8.25, 6.91, 1.22 Hz, 1H) 8.10 (d, J=8.31 Hz,1H) 8.26 (d, J=8.56 Hz, 1H).

The chloride was found to be unreactive under the alkylation protocol ofExample 184. The corresponding 6-fluoro-1,3-oxazino[5,6-c]isoquinoline(Example 226) was prepared by the method of [Uchibori, Y.; Umeno, M.;Yoshiokai, H.; Heterocycles, 1992, 34 (8), 1507-1510] cited earlier. Thereaction was not allowed to go to completion, and the crude reactionmixture was recovered as a mixture of ratio of 1:2.4 (Cl:F). Withoutfurther purification, the chloride/fluoride mixture was alkylated withthe tripeptide using the procedure of Example 184 to give 66 mg (50.0%)ofBOCNH-P3(L-t-BuGly)-P2[(4R)-(1,3-oxazino[5,6-c]isoquinoline-6-oxo)-S-proline]-P1(1R,2SVinylAcca)-CONHSO₂ Cyclopropane after preparative HPLC purification.LC/MS R_(t)-min (MNa⁺) [method D]: 3.03 (764). ¹H NMR (400 MHz, CD₃OD) δppm 1.01 (s, 9H) 1.06 (dd, J=8.07, 1.96 Hz, 2H) 1.22 (s, 10H) 1.34 (d,J=6.11 Hz, 1H) 1.42 (m, 1H) 1.86 (dd, J=8.07, 5.38 Hz, 1H) 2.23 (m, 2H)2.59 (dd, J=13.82, 6.97 Hz, 1H) 2.93 (m, 1H) 4.03 (dd, J=11.86, 3.06 Hz,1H) 4.23 (s, 1H) 4.41 (d, J=11.98 Hz, 1H) 4.50 (dd, J=9.66, 6.97 Hz, 1H)4.87 (m, 2H) 5.11 (d, J=10.52 Hz, 1H) 5.28 (d, J=17.12 Hz, 1H) 5.34 (s,2H) 5.74 (m, 2H) 7.51 (t, J=7.46 Hz, 1H) 7.70 (t, J=7.58 Hz, 1H) 7.95(d, J=8.31 Hz, 1H) 8.12 (d, J=8.31 Hz, 1H).

Preparation of 4-methoxy-3-heteroaryl and 3-azoyl isoquinoline P2*Derivatives via Suzuki and Stille Coupling Reactions

The coupling technologies shown below demonstrated the general utilitywith the bromo derivative of Example 223. It is understood that asimilar protocol is equally applicable to other combinations couplingreagents and catalysts other than boron and tin.

Example 229 Preparation of Compound 229

BOCNH-P3(L-t-BuGly)-P2[(4R)-(3-furan-3-yl-4-methoxy-isoquinolin-1-oxo)-S-proline]-P1(1R,2SVinylAcca)-CONHSO₂ Cyclopropane via a Suzuki coupling shown below:

22 mg (0.028 mmole) of Example 223 was dissolve in 1 ml of DMF, 9.4 mgof the commercial boronic acid (3 eq), 3 mg of catalyst (10% mmole) and18 mg of cesium carbonate were added. The mixture was degassed twice,and then heated up to 110° C. for 3 hours. The final product waspurified by prep-HPLC, 13.6 mg of yellow solid was obtained (64.0%).LC/MS rt-min (MH⁺): 2.85 (780) [method B]. ¹H NMR (500 MHz, CD₃OD) δ ppm1.09 (m, 11H) 1.26 (m, 12H) 1.68 (m, 1H) 2.27 (s, 1H) 2.64 (m, 2H) 2.97(m, 1H) 3.86 (s, 3H) 4.15 (d, J=10.38 Hz, 1H) 4.28 (s, 1H) 4.43 (d,J=10.99 Hz, 1H) 4.56 (m, 1H) 5.11 (m, 2H) 5.63 (m, 1H) 5.99 (s, 1H) 7.20(s, 1H) 7.51 (m, 1H) 7.61 (m, 1H) 7.75 (t, J=7.17 Hz, 1H) 8.03 (d,J=8.24 Hz, 1H) 8.16 (d, J=8.24 Hz, 1H) 8.27 (s, 1H).

Example 230 Preparation of Compound 230

BOCNH-P3(L-t-BuGly)-P2[(4R)-(3-furan-2-yl-4-methoxy-isoquinolin-1-oxo)-S-proline]-P1(1R,2SVinylAcca)-CONHSO₂ Cyclopropane was synthesized via a Stille couplingreaction shown below:

40 mg (0.05 mmole) of Example 223, 4 mg of catalyst (5% mmole) and 100μl (4 eq) of the commercial tin reagent was dissolved in 1 ml oftoluene, the mixture was degassed twice and then heated up to 90° C. forovernight. After prep HPLC separation, 19.6 mg of greenish solid wasobtained (50.0%). LC/MS rt-min (MH⁺): 2.76 (780) [method B]. ¹H NMR (400MHz, CD₃OD) δ ppm 0.94 (m, 2H) 0.98 (s, 9H) 1.09 (m, 2H) 1.25 (s, 9H)1.39 (m, 1H) 1.60 (m, 1H) 2.35 (m, 1H) 2.48 (m, 1H) 2.74 (m, 1H) 2.95(m, 1H) 3.87 (s, 3H) 4.14 (m, 1H) 4.22 (d, J=4.16 Hz, 1H) 4.41 (s, 1H)4.69 (m, 1H) 5.26 (m, 1H) 5.35 (m, 1H) 5.93 (s, 1H) 6.03 (m, 1H) 6.61(m, 1H) 7.16 (d, J=3.18 Hz, 1H) 7.50 (d, J=7.58 Hz, 1H) 7.67 (s, 1H)7.73 (t, J=7.34 Hz, 1H) 8.04 (m, 1H) 8.17 (d, J=8.31 Hz, 1H).

Example 231 Preparation of Compound 231

BOCNH-P3(L-t-BuGly)-P2[(4R)-(3-pyrazine-2-yl-4-methoxy-isoquinolin-1-oxo)-S-proline]-P1(1R,2SVinylAcca)-CONHSO₂ Cyclopropane was similarly prepared by a Stillecoupling reaction in 7.1% yield. LC/MS rt-min (MH⁺): 2.51 (792) [methodB].

¹H NMR (400 MHz, CD₃OD) δ ppm 0.98 (m, 9H) 1.11 (m, 2H) 1.19 (s, 9H)1.27 (m, 2H) 1.42 (m, 1H) 2.37 (m, 1H) 2.48 (m, 2H) 2.81 (m, 1H) 2.97(m, 1H) 3.83 (s, 3H) 4.07 (s, 1H) 4.20 (d, J=4.16 Hz, 1H) 4.54 (d,J=11.49 Hz, 1H) 4.72 (m, 1H) 5.27 (m, 1H) 5.39 (m, 1H) 5.96 (s, 1H) 6.04(m, 1H) 7.63 (s, 1H) 7.83 (s, 1H) 8.17 (s, 1H) 8.26 (s, 1H) 8.60 (d,J=2.20 Hz, 1H) 8.76 (d, J=2.20 Hz, 1H) 9.33 (s, 1H).

Example 232 Preparation of Compound 232

BOCNH-P3(L-t-BuGly)-P2[(4R)-(4-methoxy-3-thiazol-2-yl-isoquinolin-1-oxo)-S-proline]-P1(1R,2SVinylAcca)-CONHSO₂ Cyclopropane was similarly prepared by a Stillecoupling reaction in 32.2% yield. LC/MS rt-min (MH⁺): 2.42 (797) [methodB].

¹H NMR (400 MHz, CD₃OD) δ ppm 1.03 (S, 9H) 1.07 (m, 2H) 1.13 (S, 9H)1.22 (m, 2H) 1.43 (dd, J=9.78, 5.14 Hz, 1H) 1.88 (dd, J=8.07, 5.38 Hz,1H) 2.23 (q, J=8.97 Hz, 1H) 2.36 (m, 1H) 2.67 (m, 1H) 2.94 (m, 1H) 4.10(s, 3H) 4.15 (m, 1H) 4.18 (s, 1H) 4.53 (d, J=25.92 Hz, 1H) 4.59 (dd,J=10.27, 7.09 Hz, 1H) 5.12 (m, 1H) 5.29 (d, J=17.36 Hz, 1H) 5.73 (m, 1H)6.09 (s, 1H) 7.74 (t, J=7.58 Hz, 1H) 7.91 (t, J=7.70 Hz, 1H) 8.00 (d,J=3.42 Hz, 1H) 8.18 (d, J=3.18 Hz, 1H) 8.22 (d, J=8.31 Hz, 1H) 8.29 (d,J=8.31 Hz, 1H).

Example 233 Preparation of Compound 233

Following the general tripeptide alkylation procedure with thecommercial 4-chlorofuro[3,2-c]pyridine, 5.7 mg of yellow solid wasobtained (8.2%). LC/MS rt-min (MH⁺): 2.32 (674) [method B]. ¹H NMR (400MHz, CD₃OD) δ ppm 1.00 (s, 9H) 1.07 (m, 2H) 1.21 (m, 11H) 1.41 (m, 1H)1.86 (dd, J=8.07, 5.38 Hz, 1H) 2.22 (dd, J=17.61, 9.05 Hz, 2H) 2.54 (dd,J=13.69, 7.09 Hz, 1H) 2.92 (m, 1H) 4.06 (m, 1H) 4.21 (m, 1H) 4.32 (s,1H) 4.49 (m, 1H) 5.11 (dd, J=10.27, 1.47 Hz, 1H) 5.29 (dd, J=17.36, 1.22Hz, 1H) 5.74 (m, 1H) 5.81 (s, 1H) 6.83 (d, J=1.22 Hz, 1H) 7.19 (d,J=5.87 Hz, 1H) 7.76 (d, J=1.22 Hz, 1H) 7.97 (d, J=5.87 Hz, 1H).

Example 235 Preparation of Compound 235

Following the general tripeptide alkylation procedure with thecommercial 4-chlorothieno[3,2-c]pyridine, 20.0 mg of yellow solid wasobtained (28.1%). LC/MS rt-min (MH⁺): 2.50 (690) [method B]. ¹H NMR (400MHz, CD₃OD) δ ppm 1.01 (s, 9H) 1.06 (m, 2H) 1.21 (m, 11H) 1.42 (m, 1H)1.86 (dd, J=8.19, 5.50 Hz, 1H) 2.24 (m, 2H) 2.57 (dd, J=13.69, 6.85 Hz,1H) 2.93 (m, 1H) 4.05 (dd, J=11.98, 3.18 Hz, 1H) 4.22 (s, 1H) 4.39 (d,J=11.74 Hz, 1H) 4.50 (dd, J=9.90, 7.21 Hz, 1H) 5.10 (dd, J=10.39, 1.34Hz, 1H) 5.28 (d, J=17.12 Hz, 1H) 5.73 (m, 1H) 5.81 (s, 1H) 7.45 (d,J=5.62 Hz, 1H) 7.53 (m, 2H) 7.94 (d, J=5.87 Hz, 1H).

Example 236 Preparation of Compound 236

Following the general tripeptide alkylation procedure with thecommercial 3,5-dichloro-1,2,4-thiadiazole, 8.0 mg of yellow solid wasobtained (11.9%). LC/MS rt-min (MNa⁺): 2.37 (697) [method B]. ¹H NMR(400 MHz, CD₃OD) δ ppm 1.00 (s, 9H) 1.06 (m, 2H) 1.22 (m, 2H) 1.36 (s,9H) 1.42 (m, 1H) 1.86 (dd, J=8.07, 5.38 Hz, 1H) 2.25 (m, 2H) 2.60 (dd,J=14.18, 6.85 Hz, 1H) 2.92 (m, 1H) 4.03 (dd, J=12.47, 3.18 Hz, 1H) 4.17(s, 1H) 4.42 (m, 2H) 5.11 (dd, J=10.27, 1.71 Hz, 1H) 5.29 (dd, J=17.12,1.47 Hz, 1H) 5.68 (s, 1H) 5.74 (m, 1H).

Example 237 Preparation of Compound 237

BOCNH-P3(L-t-BuGly)-P2[(4R)-(quinoxaline-2-oxo)-S-proline]-P1(1R,2SVinylAcca)-CONHSO₂ Cyclopropane, Shown below

Following the general tripeptide alkylation procedure with commercial2-chloroquinoxaline, 113.0 mg of yellow solid was obtained (19.2%).LC/MS rt-min (MNa⁺): 2.48 (707) [method B]. ¹H NMR (400 MHz, CD₃OD) δppm 1.01 (s, 9H) 1.06 (m, 2H) 1.22 (m, 11H) 1.42 (m, 1H) 1.87 (dd,J=8.19, 5.50 Hz, 1H) 2.24 (m, 1H) 2.31 (m, 1H) 2.57 (dd, J=13.57, 6.97Hz, 1H) 2.93 (m, 1H) 4.09 (dd, J=11.98, 3.18 Hz, 1H) 4.17 (s, 1H) 4.38(d, J=11.74 Hz, 1H) 4.50 (dd, J=10.27, 7.09 Hz, 1H) 5.11 (dd, J=10.27,1.71 Hz, 1H) 5.29 (dd, J=17.12, 1.47 Hz, 1H) 5.74 (m, 1H) 5.87 (s, 1H)7.62 (t, J=7.46 Hz, 1H) 7.73 (t, J=7.70 Hz, 1H) 7.87 (m, 1H) 7.96 (d,J=8.31 Hz, 1H) 8.42 (s, 1H).

Example 238 Preparation of Compound 238

BOCNH-P3(L-t-BuGly)-P2[(4R)-(2-trifluoro-6-fluoroquinoline-4-oxo)-S-proline]-P1(1R,2SVinylAcca)-CONHSO₂ Cyclopropane, Shown below

Following the general tripeptide alkylation procedure with thecommercial 2-trifluoromethyl-4-chloro-6-fluoro quinoline, 17.0 mg ofyellow solid was obtained (23.2%). LC/MS rt-min (MNa⁺): 2.66 (792)[method B]. ¹H NMR (400 MHz, CD₃OD) δ ppm 1.02 (s, 9H) 1.06 (m, 2H) 1.17(s, 9H) 1.23 (m, 2H) 1.42 (m, 1H) 1.86 (dd, J=8.07, 5.38 Hz, 1H) 2.21(q, J=8.64 Hz, 1H) 2.32 (m, 1H) 2.63 (dd, J=13.94, 6.85 Hz, 1H) 2.93 (m,1H) 4.08 (m, 1H) 4.18 (s, 1H) 4.53 (m, 2H) 5.10 (m, 1H) 5.27 (d, J=17.12Hz, 1H) 5.58 (s, 1H) 5.72 (m, 1H) 7.39 (s, 1H) 7.65 (m, 1H) 7.83 (dd,J=9.29, 2.69 Hz, 1H) 8.12 (dd, J=9.29, 5.14 Hz, 1H).

Example 239 Preparation of Compound 239

BOCNH-P3(L-t-BuGly)-P2[(4R)-(6-fluoroquinoline-4-oxo)-S-proline]-P1(1R,2SVinylAcca)-CONHSO₂ Cyclopropane, Shown below

Following the general tripeptide alkylation procedure with thecommercial 4-chloro-6-fluoro quinoline, 26.0 mg of yellow solid wasobtained (39.0%). LC/MS rt-min (MH⁺): 1.98 (702) [method B]. ¹H NMR (400MHz, CD₃OD) δ ppm 1.04 (m, 11H) 1.14 (s, 9H) 1.23 (m, 2H) 1.42 (m, 1H)1.87 (dd, J=8.07, 5.38 Hz, 1H) 2.23 (q, J=8.80 Hz, 1H) 2.41 (m, 1H) 2.75(dd, J=14.43, 6.85 Hz, 1H) 2.93 (m, 1H) 4.09 (s, 1H) 4.12 (d, J=2.69 Hz,1H) 4.61 (m, 2H) 5.11 (dd, J=10.39, 1.59 Hz, 1H) 5.28 (dd, J=17.24, 1.34Hz, 1H) 5.70 (m, 1H) 5.75 (s, 1H) 7.62 (d, J=6.60 Hz, 1H) 7.93 (m, 1H)8.06 (dd, J=8.68, 2.57 Hz, 1H) 8.20 (dd, J=9.29, 4.40 Hz, 1H) 9.06 (d,J=6.60 Hz, 1H). A small amount of the by-product due to F-displacementwas also isolated from the same reaction and was separated bypreparative HPLC.

Example 240 Isolation of Compound 240

BOCNH-P3(L-t-BuGly)-P2[(4R)-(4-chloroquinoline-6-oxo)-S-proline]-P1(1R,2SVinylAcca)-CONHSO₂ Cyclopropane, Shown below

By-product, 8.0 mg of yellow solid was obtained (11.7%). LC/MS rt-min(MNa⁺): 2.240 (740) [method B]. ¹H NMR (400 MHz, CD₃OD) δ ppm 1.00 (s,9H) 1.06 (m, 2H) 1.23 (m, 11H) 1.42 (m, 1H) 1.86 (dd, J=8.19, 5.50 Hz,1H) 2.22 (m, 1H) 2.29 (m, 1H) 2.55 (m, 1H) 2.92 (m, 1H) 4.09 (m, 1H)4.21 (s, 1H) 4.30 (m, 1H) 4.46 (dd, J=10.27, 6.85 Hz, 1H) 5.11 (dd,J=10.27, 1.47 Hz, 1H) 5.28 (dd, J=17.36, 1.47 Hz, 1H) 5.43 (s, 1H) 5.74(m, 1H) 7.60 (dd, J=9.29, 2.45 Hz, 1H) 7.64 (d, J=2.45 Hz, 1H) 7.81 (d,J=4.89 Hz, 1H) 8.06 (d, J=9.29 Hz, 1H) 8.72 (d, J=5.14 Hz, 1H).

Example 241 Preparation of Compound 241

BOCNH-P3(L-t-BuGly)-P2[(4R)-(8-fluoroquinoline-4-oxo)-S-proline]-P1(1R,2SVinylAcca)-CONHSO₂ Cyclopropane, Shown below

Following the general tripeptide alkylation procedure with thecommercial 4-chloro-8-fluoro quinoline, 10.3 mg of yellow solid wasobtained (14.7%). LC/MS rt-min (MH⁺): 1.95 (702) [method B]. ¹H NMR (400MHz, CD₃OD) δ ppm 0.98 (s, 9H) 1.06 (m, 2H) 1.14 (s, 9H) 1.22 (m, 2H)1.42 (m, 1H) 1.87 (dd, J=8.07, 5.62 Hz, 1H) 2.22 (q, J=8.72 Hz, 1H) 2.41(m, 1H) 2.74 (dd, J=14.06, 6.97 Hz, 1H) 2.93 (m, 1H) 4.11 (m, 2H) 4.57(dd, J=10.39, 6.97 Hz, 1H) 4.66 (d, J=12.23 Hz, 1H) 5.11 (dd, J=10.27,1.22 Hz, 1H) 5.28 (d, J=17.12 Hz, 1H) 5.71 (m, 2H) 7.59 (d, J=6.36 Hz,1H) 7.75 (m, 1H) 7.86 (m, 1H) 8.23 (d, J=8.56 Hz, 1H) 9.02 (d, J=6.36Hz, 1H). During the preparative HPLC purification, a by-product was alsoisolated. The 4-chloroquinoline-8-oxo-quinoline derivative was formed asa result of displacement of the fluorine atom instead of the chlorineleaving group.

Example 242 Isolation of Compound 242

BOCNH-P3(L-t-BuGly)-P2[(4R)-(4-chloroquinoline-8-oxo)-S-proline]-P1(1R,2SVinylAcca)-CONHSO₂ Cyclopropane, Shown below

Thus the by-product, 9.0 mg of yellow solid was obtained (13.2%). LC/MSrt-min (MH⁺): 2.37 (718) [method B]. ¹H NMR (400 MHz, CD₃OD) δ ppm 1.00(s, 9H) 1.06 (m, 2H) 1.12 (s, 9H) 1.23 (m, 2H) 1.43 (dd, J=9.41, 5.50Hz, 1H) 1.87 (dd, J=8.19, 5.50 Hz, 1H) 2.25 (m, 1H) 2.35 (m, 1H) 2.67(dd, J=13.94, 7.09 Hz, 1H) 2.93 (m, 1H) 4.10 (m, 1H) 4.13 (s, 1H) 4.43(d, J=11.98 Hz, 1H) 4.65 (dd, J=10.03, 7.09 Hz, 1H) 5.12 (dd, J=10.27,1.47 Hz, 1H) 5.30 (dd, J=17.12, 1.22 Hz, 1H) 5.51 (s, 1H) 5.75 (m, 1H)7.61 (d, J=7.83 Hz, 1H) 7.88 (t, J=8.19 Hz, 1H) 8.04 (m, 2H) 8.91 (d,J=5.38 Hz, 1H).

Example 243 Preparation of Compound 243

BOCNH-P3(L-t-BuGly)-P2[(4R)-(3-hydroxyquinoxaline-2-oxo)-S-proline]-P1(1R,2SVinylAcca)-CONHSO₂ Cyclopropane, Shown below

Following the general tripeptide alkylation procedure with commercial2,3-dichloroquinoxaline, the mono alkylation product was spontaneouslyhydrolyzed to give 8.0 mg of pale yellow solid (11.4%). LC/MS rt-min(MNa⁺): 2.42 (723) [method B]. ¹H NMR (400 MHz, CD₃OD) δ ppm 0.99 (s,9H) 1.05 (m, 2H) 1.24 (s, 9H) 1.40 (m, 3H) 1.86 (m, 1H) 2.24 (m, 2H)2.53 (m, 1H) 2.92 (m, 1H) 4.06 (m, 1H) 4.16 (s, 1H) 4.40 (m, 1H) 4.55(dd, J=10.39, 6.97 Hz, 1H) 5.11 (m, 1H) 5.29 (m, 1H) 5.73 (m, 1H) 5.78(s, 1H) 7.15 (s, 1H) 7.26 (m, 2H) 7.36 (t, J=7.83 Hz, 1H) 7.61 (d,J=8.07 Hz, 1H).

Example 244 Preparation of Compound 244

Using a combination of Pd^(o) coupling scheme and a step by stepprocedure starting from 6-bromo-1-chloro isoquinoline,BOCNH-P3(L-t-BuGly)-P2[(4R)-(6-carboxylic aciddimethylamideisoquinoline-1-oxo)-S-proline]-P1(1R,2S VinylAcca)-CONHSO₂Cyclopropane, was prepared.

LC/MS rt-min (MNa⁺): 2.34 (777) [method B]. ¹H NMR (400 MHz, CD₃OD) δppm 0.98 (m, 11H) 1.23 (m, 11H) 1.35 (m, 1H) 1.91 (m, 1H) 2.29 (m, 2H)2.47 (m, 1H) 2.58 (m, 1H) 2.97 (s, 3H) 3.11 (s, 3H) 4.09 (m, 1H) 4.24(s, 1H) 4.44 (m, 1H) 4.61 (m, 1H) 5.16 (m, 2H) 5.57 (m, 1H) 5.90 (s, 1H)7.38 (d, J=5.87 Hz, 1H) 7.50 (d, J=8.07 Hz, 1H) 7.86 (s, 1H) 8.03 (d,J=5.87 Hz, 1H) 8.27 (d, J=8.56 Hz, 1H),

Example 245 Preparation of Compound 245

During one of the Pd⁰ catalyzed Stille coupling preparations (Example230), a side product was isolated as a minor product which wassubsequently identified as:

BOCNH-P3(L-t-BuGly)-P2[(4R)-(3-chloro-4-methoxyisoquinoline-1-oxo)-S-proline]-P1(1R,2SVinylAcca)-CONHSO₂ Cyclopropane, shown below

LC/MS rt-min (MNa⁺): 2.62 (770) [method B]. ¹H NMR (400 MHz, CD₃OD) δppm 1.01 (s, 9H) 1.08 (m, 2H) 1.18 (s, 9H) 1.27 (m, 2H) 1.37 (m, 1H)1.62 (m, 1H) 2.36 (m, 2H) 2.73 (m, 1H) 2.97 (m, 1H) 3.92 (s, 3H) 4.02(m, 1H) 4.18 (s, 1H) 4.48 (m, 1H) 4.66 (m, 1H) 5.30 (m, 2H) 5.78 (s, 1H)6.04 (m, 1H) 7.53 (t, J=7.70 Hz, 1H) 7.77 (t, J=7.58 Hz, 1H) 8.00 (d,J=8.56 Hz, 1H) 8.19 (d, J=8.07 Hz, 1H).

Section F Example 250 Preparation of Compound 250

Step 1:

A solution of 3-phenyl-but-2-enoic acid (16.2 g), diphenylphosphorylazide (27.5 g), and triethylamine (10.1 g) in benzene (100 mL) wasstirred for 1 h. After filtration through a silica gel plug washing withbenzene and concentration, the residue was dissolved in diphenylmethane(80 mL) and refluxed for 3 h. After cooling to rt, solids were collectedthrough a plug washing with benzene and dried to give 10 g (63%) of thedesired product as a solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 2.30 (s, 3H),7.00 (s, 1H), 7.54 (m, 1H), 7.77 (m, 2H), 8.33 (d, J=7.34 Hz, 1H).

Step 2

A solution of 4-methyl-2H-isoquinolin-1-one (4.8 g) in POCl₃ (50 mL) wasrefluxed for 3 h. After cooling and concentration, the residue was basedwith 5 N NaOH and extracted with CH₂Cl₂. The organic layer was washedwith brine and dried over MgSO₄. After concentration, purification byflash chromatography of Biotage with 5% ethyl acetate in hexanes gave4.8 g (90%) of the desired product as a solid. ¹H NMR (400 MHz, CDCl₃) δppm 2.59 (s, 3H), 7.68 (t, J=7.70 Hz, 1H), 7.78 (m, 1H), 7.94 (d, J=8.31Hz, 1H), 8.11 (s, 1H), 8.35 (d, J=8.31 Hz, 1H).

Step 3:

A solution of Boc-Hyp-OH (231 mg) and tert-BuOK (336 mg) in DMSO (10 mL)was stirred for 0.5 h. To the solution was added1-chloro-4-methyl-isoquinoline (178 mg) and the resulting mixture wasstirred for 1 day. The reaction was quenched with 5% citric acid andextracted with ethyl acetate. The organic layer was washed with brineand dried over MgSO₄. Concentration gave 350 mg (94%) of the desiredproduct as a solid which was used in the next step without furtherpurification. ¹H NMR (400 MHz, CD₃OD) δ ppm 1.39, 1.43 (2s, 9H,rotamers), 2.40 (dd, J=17.97, 4.52 Hz, 1H), 2.48 (s, 3H), 2.68 (m, 1H),3.84 (m, 2H), 4.46 (m, 1H), 5.71 (s, 1H), 7.58 (t, J=7.70 Hz, 1H), 7.75(m, 2H), 7.91 (d, J=8.31 Hz, 1H), 8.19 (m, 1H); MS: (M+Na)⁺ 396.

Step 4:

A solution of4-(4-methyl-isoquinolin-1-yloxy)-pyrrolidine-1,2-dicarboxylic acid1-tert-butyl ester) (74 mg), cyclopropanesulfonic acid(1(R)-amino-2(S)-vinyl-cyclopropanecarbonyl)-amide hydrochloride (59mg), PyBOP (114 mg) and i-Pr₂NEt (0.2 mL) in CH₂Cl₂ (2 mL) was stirredfor 2 h. Purification by flash chromatograph of Biotage with 5% MeOH inethyl acetate gave 105 mg (90%) of the desired product. ¹H NMR (400 MHz,Methanol-D4) δ ppm 1.18 (m, 5H), 1.39 (s, 9H), 1.87 (dd, J=8.2, 5.3 Hz,1H), 2.28 (m, 2H), 2.54 (m, 4H), 2.95 (m, 1H), 3.86 (m, 2H), 4.40 (dd,J=9.8, 6.9 Hz, 1H), 5.12 (d, J=10.5 Hz, 1H), 5.31 (d, J=17.6 Hz, 1H),5.79 (m, 2H), 7.60 (t, J=7.5 Hz, 1H), 7.78 (m, 2H), 7.93 (d, J=8.3 Hz,1H), 8.20 (d, J=8.1 Hz, 1H); MS: (M+Na)⁺ 607.

Step 5:

A solution of2-(1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-(4-methyl-isoquinolin-1-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester

(100 mg) and TFA (3 mL) in CH₂Cl₂ (3 mL) was stirred for 1 h. Afterconcentration, the residue was dissolved in CH₂Cl₂ (2 mL), andBoc-L-tert-leucine (40 mg), PyBOP (104 mg) and i-Pr₂NEt (0.2 mL) wasadded. The mixture was stirred for 1 h. After work-up, purification PrepHPLC gave 60 mg (52%) of the desired product compound 250 as a solid. ¹HNMR (400 MHz, CD₃OD) δ ppm 1.04 (m, 12H), 1.26 (m, 10H), 1.44 (dd,J=9.5, 5.1 Hz, 1H), 1.88 (dd, J=8.1, 5.4 Hz, 1H), 2.26 (m, 2H), 2.49 (s,3H), 2.62 (dd, J=13.7, 7.1 Hz, 1H), 2.94 (m, 1H), 4.06 (dd, J=12.0, 3.4Hz, 1H), 4.25 (m, 1H), 4.45 (d, J=11.3 Hz, 1H), 4.53 (dd, J=10.3, 6.6Hz, 1H), 5.12 (d, J=10.0 Hz, 1H), 5.29 (d, J=17.1 Hz, 1H), 5.77 (m, 2H),6.63 (d, J=8.6 Hz, 1H), 7.53 (t, J=7.8 Hz, 1H), 7.76 (t, J=8.1 Hz, 1H),7.80 (s, 1H), 7.91 (d, J=8.1 Hz, 1H), 8.22 (d, J=8.3 Hz, 1H); MS:(M+Na)⁺ 720.

Example 251 Preparation of Compound 251

Compound 251 was prepared by following Scheme 1 of Example 250 exceptthat 3-methoxy-3-phenyl-acrylic acid was used in place of3-phenyl-but-2-enoic acid in step 1.

Step 1:

Modifications: 15 g 3-methoxy-3-phenyl-acrylic acid used, 250 mg productobtained (2% yield).

Product:

¹H NMR (400 MHz, CD₃COCD₃) δ ppm 3.85 (s, 3H), 6.96 (s, 1H), 7.54 (m,1H), 7.71 (m, 1H), 7.86 (d, J=8.07 Hz, 1H), 8.31 (d, J=8.07 Hz, 1H).

Step 2:

Modifications: 200 mg 4-methoxy-2H-isoquinolin-1-one used, 150 mgproduct obtained (68% yield).

Product:

¹H NMR (400 MHz, CDCl₃) δ ppm 4.05 (s, 2H), 7.71 (m, 1H), 7.72 (m, 2H),7.80 (s, 1H), 8.23 (dd, J=18.71, 7.70 Hz, 2H).

Step 3:

Modifications: 122 mg 1-chloro-4-methoxy-isoquinoline used, 218 mgproduct obtained (89% yield).

Product:

MS: (M+Na)⁺ 411.

Step 4:

Modifications: 194 mg4-(4-methoxy-isoquinolin-1-yloxy)-pyrrolidine-1,2-dicarboxylic acid1-tert-butyl ester used, 298 mg product obtained (99% yield).

Product:

¹H NMR (400 MHz, CD₃OD) δ ppm 1.17 (m, 5H), 1.42 (s, 9H), 1.87 (dd,J=8.2, 5.5 Hz, 1H), 2.27 (m, 2H), 2.54 (dd, J=13.3, 6.2 Hz, 1H), 2.95(m, 1H), 3.85 (m, 2H), 4.00 (s, 3H), 4.39 (dd, J=9.8, 6.9 Hz, 1H), 5.12(d, J=10.5 Hz, 1H), 5.31 (d, J=17.1 Hz, 1H), 5.76 (m, 2H), 7.52 (s, 1H),7.62 (t, J=7.6 Hz, 1H), 7.74 (t, J=7.2 Hz, 1H), 8.12 (t, J=8.3 Hz, 2H).

Step 5:

Modifications: 190 mg2-(1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-(4-methoxy-isoquinolin-1-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester used, 270 mg product obtained (51% yield).

Product:

Data: ¹H NMR (500 MHz, CD₃OD) δ ppm 1.06 (m, 12H), 1.26 (m, 10H), 1.43(dd, J=8.6, 4.6 Hz, 1H), 1.88 (dd, J=7.9, 5.5 Hz, 1H), 2.24 (m, 2H),2.61 (dd, J=13.6, 6.9 Hz, 1H), 2.94 (m, 1H), 4.00 (s, 3H), 4.06 (dd,J=11.3, 3.1 Hz, 1H), 4.25 (d, J=8.9 Hz, 1H), 4.43 (d, J=11.3 Hz, 1H),4.52 (m, 1H), 5.12 (d, J=10.1 Hz, 1H), 5.29 (d, J=17.1 Hz, 1H), 5.75 (m,2H), 6.60 (d, J=8.6 Hz, 1H), 7.55 (m, 2H), 7.71 (t, J=7.3 Hz, 1H), 8.09(d, J=8.2 Hz, 1H), 8.14 (d, J=8.2 Hz, 1H); MS: (M+Na)⁺ 736.

Example 252 Preparation of Compound 252

Compound 252 was prepared by following Scheme 1 of Example 250 except2-methylcinnamic acid was used in place of 3-phenyl-but-2-enoic acid instep 1.

Step 1:

Modifications: 20 g 2-methylcinnamic acid used, 14.3 g product obtained(72% yield)

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 2.54 (s, 1H), 6.69 (d, J=7.3 Hz,1H), 7.23 (d, J=7.3 Hz, 1H), 7.39 (t, J=7.8 Hz, 1H), 7.50 (d, J=7.1 Hz,1H), 8.30 (d, J=8.1 Hz, 1H), 11.62 (s, 1H); MS: (M+H)⁺ 160.

Step 2:

Modifications: 14.4 g 5-methyl-2H-isoquinolin-1-one used, 10.6 g productobtained (66% yield).

Product:

Data: ¹H NMR (400 MHz, CDCl₃) δ ppm 2.67 (s, 3H), 7.55 (m, 2H), 7.70(dd, J=5.9, 1.0 Hz, 1H), 8.19 (m, 1H), 8.28 (d, J=5.9 Hz, 1H); MS:(M+H)⁺ 178.

Step 3:

Modifications: 533 mg 1-chloro-5-methyl-isoquinoline used, 1116 mgproduct obtained (100% yield).

Product:

Data: MS: (M+H)⁺ 373.

Step 4:

Modifications: 372 mg4-(5-methyl-isoquinolin-1-yloxy)-pyrrolidine-1,2-dicarboxylic acid1-tert-butyl ester used, 551 mg product obtained (94% yield).

Product:

Data: MS: (M+Na)⁺ 607.

Step 5:

Modifications: 551 mg2-(1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-(5-methyl-isoquinolin-1-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester used, 274 mg product obtained (44% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 1.00 (m, 12H), 1.23 (m, 10H), 1.44(m, 1H), 1.87 (dd, J=8.1, 5.4 Hz, 1H), 2.26 (m, 2H), 2.62 (m, 4H), 2.94(m, 1H), 4.07 (dd, J=11.9, 3.3 Hz, 1H), 4.25 (d, J=9.5 Hz, 1H), 4.46 (d,J=11.5 Hz, 1H), 4.53 (dd, J=10.3, 7.1 Hz, 1H), 5.12 (d, J=10.5 Hz, 1H),5.29 (d, J=16.9 Hz, 1H), 5.75 (m, 1H), 5.86 (s, 1H), 6.62 (d, J=9.3 Hz,1H), 7.39 (t, J=7.7 Hz, 1H), 7.44 (d, J=5.9 Hz, 1H), 7.53 (d, J=7.1 Hz,1H), 8.00 (d, J=6.1 Hz, 1H), 8.06 (d, J=8.3 Hz, 1H); MS: (M+H)⁺ 698.

Example 253 Preparation of Compound 253

Compound 253 was prepared by following Scheme 1 of Example 250 except2-methoxy cinnamic acid was used in place of 3-phenyl-but-2-enoic acidin step 1.

Step 1:

Modifications: 10 g 2-methoxy cinnamic acid used, 5.3 g product obtained(53% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 3.95 (s, 3H), 6.94 (d, J=7.3 Hz,1H), 7.08 (d, J=8.1 Hz, 1H), 7.14 (d, J=7.3 Hz, 1H), 7.43 (t, J=8.1 Hz,1H), 7.99 (d, J=8.1 Hz, 1H), 10.92 (s, 1H); MS: (M+H)⁺ 176.

Step 2:

Modifications: 5.3 g 5-methoxy-2H-isoquinolin-1-one used, 5.38 g productobtained (92% yield).

Product:

Data: ¹H NMR (400 MHz, CDCl₃) δ ppm 4.01 (s, 3H), 7.04 (d, J=7.8 Hz,1H), 7.57 (t, J=8.1 Hz, 1H), 7.88 (d, J=8.6 Hz, 1H), 7.97 (d, J=5.9 Hz,1H), 8.25 (d, J=5.9 Hz, 1H); MS: (M+H)⁺ 194.

Step 3:

Modifications: 581 mg 1-chloro-5-methoxy-isoquinoline used, 1163 mgproduct obtained (100% yield).

Product:

Data: MS: (M+H)⁺ 389.

Step 4:

Modifications: 117 mg4-(5-methoxy-isoquinolin-1-yloxy)-pyrrolidine-1,2-dicarboxylic acid1-tert-butyl ester used, 180 mg product obtained (100% yield).

Product:

Data: MS: (M+H)⁺ 601.

Step 5:

Modifications: 177 mg2-(1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-(5-methoxy-isoquinolin-1-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester used, 63 mg product obtained (44% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 1.00 (m, 12H), 1.21 (m, 10H), 1.38(m, 1H), 1.82 (dd, J=8.1, 5.6 Hz, 1H), 2.20 (m, 2H), 2.56 (dd, J=13.6,6.7 Hz, 1H), 2.88 (m, 1H), 3.08 (m, 2H), 3.93 (s, 3H), 4.01 (dd, J=11.9,3.3 Hz, 1H), 4.20 (d, J=9.1 Hz, 1H), 4.39 (d, J=12.2 Hz, 1H), 4.47 (dd,J=9.7, 7.0 Hz, 1H), 5.06 (d, J=10.0 Hz, 1H), 5.23 (d, J=16.9 Hz, 1H),5.70 (m, 1H), 5.79 (s, 1H), 6.55 (d, J=9.5 Hz, 1H), 7.08 (d, J=7.6 Hz,1H), 7.37 (t, J=8.0 Hz, 1H), 7.54 (d, J=5.9 Hz, 1H), 7.68 (d, J=8.3 Hz,1H), 7.89 (d, J=5.9 Hz, 1H); MS: (M+H)⁺ 714.

Example 254 Preparation of Compound 254

Compound 254 was prepared by following Scheme 1 of Example 250 exceptthat 2-chlorocinnamic acid was used in place of 3-phenyl-but-2-enoicacid in step 1.

Step 1:

Modifications: 25 g 2-chlorocinnamic acid used, 14.6 g product obtained(59% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 7.22 (d, J=7.3 Hz, 1H), 7.42 (t,J=7.8 Hz, 1H), 7.73 (d, J=7.8 Hz, 1H), 8.34 (d, J=8.1 Hz, 1H), 10.61 (s,1H); MS: (M+H)⁺ 180.

Step 2:

Modifications: 14.2 g 5-chloro-2H-isoquinolin-1-one used, 8.28 g productobtained (53% yield).

Product:

Data: ¹H NMR (400 MHz, CDCl₃) δ ppm 7.60 (dd, J=8.6, 7.6 Hz, 1H), 7.83(m, 1H), 8.00 (d, J=5.9 Hz, 1H), 8.29 (dt, J=8.9, 1.0 Hz, 1H), 8.38 (d,J=5.9 Hz, 1H); MS: (M+H)⁺ 198.

Step 3:

Modifications: 594 mg 1,5-dichloro-isoquinoline used, 1174 mg productobtained (100% yield).

Product:

Data: MS: (M+H)⁺ 393.

Step 4:

Modifications: 118 mg4-(5-chloro-isoquinolin-1-yloxy)-pyrrolidine-1,2-dicarboxylic acid1-tert-butyl ester used, 154 mg product obtained (85% yield).

Product:

Data: MS: (M+H)⁺ 605.

Step 5:

Modifications: 150 mg2-(1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-(5-chloro-isoquinolin-1-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester used, 91 mg product obtained (51% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 0.97 (m, 12H), 1.17 (m, 10H), 1.38(dd, J=9.4, 5.3 Hz, 1H), 1.82 (dd, J=8.0, 5.5 Hz, 1H), 2.21 (m, 2H),2.58 (dd, J=13.8, 7.0 Hz, 1H), 2.88 (m, 1H), 4.01 (dd, J=11.9, 2.8 Hz,1H, 4.16 (d, J=9.3 Hz, 1H), 4.47 (m, 2H), 5.06 (d, J=10.3 Hz, 1H), 5.24(d, J=16.9 Hz, 1H), 5.70 (m, 1H), 5.82 (s, 1H), 6.52 (d, J=9.3 Hz, 1H),7.42 (t, J=8.0 Hz, 1H), 7.57 (d, J=6.1 Hz, 1H), 7.76 (d, J=7.6 Hz, 1H),8.05 (d, J=6.1 Hz, 1H), 8.13 (d, J=8.3 Hz, 1H); MS: (M+H)⁺ 718.

Example 255 Preparation of Compound 255

Compound 255 was prepared by following Scheme 1 of Example 250 exceptthat 2-fluorocinnamic acid was used in place of 3-phenyl-but-2-enoicacid in step 1.

Step 1:

Modifications: 16.6 g 2-fluorocinnamic acid used, 8.55 g productobtained (51% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃COCD₃) δ ppm 6.62 (d, J=7.3 Hz, 1H), 7.32 (d,J=7.3 Hz, 1H), 7.47 (m, 2H), 8.09 (m, 1H).

Step 2:

Modifications: 8.4 g 5-fluoro-2H-isoquinolin-1-one used, 7.5 g productobtained (80% yield).

Product:

Data: ¹H NMR (400 MHz, CDCl₃) δ ppm 7.43 (ddd, J=9.7, 7.8, 0.9 Hz, 1H),7.62 (td, J=8.2, 5.4 Hz, 1H), 7.84 (d, J=5.6 Hz, 1H), 8.14 (d, J=8.6 Hz,1H), 8.33 (d, J=5.9 Hz, 1H); MS: (M+H)⁺ 182.

Step 3:

Modifications: 203 mg 1-chloro-5-fluoro-isoquinoline used, 384 mgproduct obtained (90% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃SOCD₃) δ ppm 1.34, 1.36 (2s, 9H, rotamers),2.35 (m, 1H), 2.61 (m, 1H), 3.65 (d, J=12.23 Hz, 1H), 3.80 (m, 1H), 4.35(m, 1H), 5.70 (s, 1H), 7.48 (d, J=6.11 Hz, 1H), 7.63 (m, 2H), 7.99 (m,1H), 8.10 (d, J=5.87 Hz, 1H); MS: (M+Na)⁺ 399.

Step 4:

Modifications: 76 mg4-(5-fluoro-isoquinolin-1-yloxy)-pyrrolidine-1,2-dicarboxylic acid1-tert-butyl ester used, 116 mg product obtained (99% yield).

Product:

Step 5:

Modifications: 110 mg2-(1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-(5-fluoro-isoquinolin-1-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester used, 39 mg product obtained (30% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 1.05 (m, 12H), 1.25 (m, 10H), 1.44(dd, J=9.5, 5.4 Hz, 1H), 1.88 (dd, J=8.1, 5.4 Hz, 1H), 2.28 (m, 2H),2.63 (dd, J=13.8, 7.0 Hz, 1H), 2.94 (m, 1H), 4.07 (dd, J=11.9, 3.1 Hz,1H), 4.23 (d, J=9.3 Hz, 1H), 4.52 (m, 2H), 5.12 (dd, J=10.3, 1.5 Hz,1H), 5.29 (d, J=17.4 Hz, 1H), 5.75 (m, 1H), 5.89 (s, 1H), 6.59 (d, J=9.1Hz, 1H), 7.47 (m, 3H), 8.02 (d, J=8.1 Hz, 1H), 8.06 (d, J=6.1 Hz, 1H);MS: (M+Na)⁺ 724.

Example 256 Preparation of Compound 256

Compound 256 was prepared by following Scheme 1 of Example 250 except2-difluormethoxycinnamic acid was used in place of 3-phenyl-but-2-enoicacid in step 1.

Step 1:

Modifications: 10.7 g 2-difluormethoxycinnamic acid used, 2 g productobtained (18% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃SOCD₃) δ ppm 6.06 (m, 2H), 6.42 (m, 2H), 6.71(s, 2H), 7.35 (s, 1H); MS: (M+H)⁺ 212.

Step 2:

Modifications: 300 mg 5-difluoromethoxy-2H-isoquinolin-1-one used, 300mg product obtained (92% yield).

Product:

Data: ¹H NMR (400 MHz, CDCl₃) δ ppm 6.70 (t, J=72.87 Hz, 1H), 7.48 (m,1H), 7.64 (m, 1H), 7.92 (d, J=5.87 Hz, 1H), 8.21 (d, J=8.56 Hz, 1H),8.35 (d, J=5.62 Hz, 1H).

Step 3:

Modifications: 230 mg 1-chloro-5-difluoromethoxy-isoquinoline used, 360mg product obtained (96% yield).

Product:

Step 4:

Modifications: 37 mg4-(5-hydroxy-isoquinolin-1-yloxy)-pyrrolidine-1,2-dicarboxylic acid1-tert-butyl ester used, 57 mg product obtained (99% yield).

Product:

Step 5:

Modifications: 57 mg2-(1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-(5-hydroxy-isoquinolin-1-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester used, 10 mg product obtained (15% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 0.93 (m, 4H), 1.13 (s, 9H), 1.31 (m,1H), 1.49 (s, 9H), 1.89 (dd, J=7.8, 5.4 Hz, 1H), 2.16 (q, J=8.8 Hz, 1H),2.40 (m, 1H), 2.81 (m, 1H), 2.90 (m, 1H), 3.76 (m, 2H), 4.30 (m, 1H),4.59 (dd, J=10.2, 7.7 Hz, 1H), 5.07 (dd, J=10.3, 1.7 Hz, 1H), 5.26 (dd,J=17.2, 1.3 Hz, 1H), 5.77 (dt, J=17.2, 9.6 Hz, 1H), 5.93 (s, 1H), 7.24(d, J=8.6 Hz, 1H), 7.51 (m, 2H), 7.63 (t, J=8.0 Hz, 1H), 7.98 (d, J=6.1Hz, 1H), 8.24 (d, J=8.3 Hz, 1H); MS: (M+H)⁺ 700.

Example 257 Preparation of Compound 257

Compound 257 was prepared by following Scheme 1 of Example 250 except4-fluorocinnamic acid was used in place of 3-phenyl-but-2-enoic acid instep 1.

Step 1:

Modifications: 16.6 g 4-fluorocinnamic acid used, 8.2 g product obtained(49% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃COCD₃) δ ppm 6.57 (d, J=7.09 Hz, 1H), 7.21 (d,J=7.09 Hz, 1H), 7.50 (m, 1H), 7.72 (dd, J=8.68, 5.26 Hz, 1H), 7.90 (dd,J=9.54, 2.93 Hz, 1H).

Step 2:

Modifications: 8.15 g 7-fluoro-2H-isoquinolin-1-one used, 7.6 g productobtained (84% yield).

Product:

Data: ¹H NMR (400 MHz, CDCl₃) δ ppm 7.52 (td, J=8.6, 2.6 Hz, 1H), 7.59(d, J=5.6 Hz, 1H), 7.86 (dd, J=9.1, 5.4 Hz, 1H), 7.95 (dd, J=9.5, 2.5Hz, 1H), 8.26 (d, J=5.6 Hz, 1H); MS: (M+H)⁺ 182.

Step 3:

Modifications: 191 mg 1-chloro-7-fluoro-isoquinoline used, 350 mgproduct obtained (93% yield).

Product:

Data: MS: (M+Na)⁺ 399.

Step 4:

Modifications: 75 mg4-(7-fluoro-isoquinolin-1-yloxy)-pyrrolidine-1,2-dicarboxylic acid1-tert-butyl ester used, 100 mg product obtained (85% yield)

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 1.16 (m, 4H), 1.41 (m, 10H), 1.88(dd, J=8.1, 5.4 Hz, 1H), 2.28 (m, 2H), 2.56 (m, 1H), 2.94 (m, 1H), 3.87(m, 2H), 4.41 (dd, J=9.7, 7.0 Hz, 1H), 5.12 (d, J=10.8 Hz, 1H), 5.31 (d,J=17.1 Hz, 1H), 5.78 (m, 2H), 7.36 (d, J=5.9 Hz, 1H), 7.54 (m, 1H), 7.78(dd, J=9.3, 2.5 Hz, 1H), 7.90 (dd, J=9.1, 5.1 Hz, 1H), 7.96 (d, J=5.9Hz, 1H); MS: (M+Na)⁺ 611.

Step 5:

Modifications: 95 mg2-(1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-(7-fluoro-isoquinolin-1-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester used, 55 mg product obtained (44% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 1.05 (m, 12H), 1.22 (m, 10H), 1.44(dd, J=9.3, 5.4 Hz, 1H), 1.88 (dd, J=8.2, 5.5 Hz, 1H), 2.27 (m, 2H),2.63 (dd, J=13.8, 7.0 Hz, 1H), 2.94 (m, 1H), 4.07 (dd, J=11.5, 3.2 Hz,1H), 4.22 (d, J=9.5 Hz, 1H), 4.47 (d, J=11.7 Hz, 1H), 4.55 (dd, J=10.6,7.5 Hz, 1H), 5.12 (d, J=10.3 Hz, 1H), 5.29 (d, J=17.1 Hz, 1H), 5.75 (m,1H), 5.87 (s, 1H), 6.61 (d, J=9.5 Hz, 1H), 7.36 (d, J=5.9 Hz, 1H), 7.52(td, J=8.9, 2.5 Hz, 1H), 7.79 (dd, J=9.4, 2.6 Hz, 1H), 7.88 (dd, J=8.7,5.5 Hz, 1H), 7.96 (d, J=5.9 Hz, 1H); MS: (M+Na)⁺ 724.

Example 258 Preparation of Compound 258

Compound 258 was prepared by following Scheme 1 of Example 250 exceptthat 4-chlorocinnamic acid was used in place of 3-phenyl-but-2-enoicacid in step 1.

Step 1:

Modifications: 9.13 g 4-chlorocinnamic acid used, 4 g product obtained(44% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃SOCD₃) δ ppm 6.58 (d, J=7.1 Hz, 1H), 7.20 (dd,J=7.1, 5.9 Hz, 1H), 7.72 (m, 2H), 8.10 (m, 1H).

Step 2:

Modifications: 3.5 g 7-chloro-2H-isoquinolin-1-one used, 2.8 g productobtained (72% yield).

Product:

Data: ¹H NMR (500 MHz, CDCl₃) δ ppm 7.59 (d, J=5.5 Hz, 1H), 7.69 (dd,J=8.9, 2.1 Hz, 1H), 7.80 (d, J=8.6 Hz, 1H), 8.29 (d, J=5.5 Hz, 1H), 8.34(s, 1H); MS: (M+H)⁺ 198.

Step 3:

Modifications: 208 mg 1,7-dichloro-isoquinoline used, 350 mg productobtained (89% yield).

Product:

Data: MS: (M+Na)⁴¹⁵.

Step 4:

Modifications: 79 mg4-(7-chloro-isoquinolin-1-yloxy)-pyrrolidine-1,2-dicarboxylic acid1-tert-butyl ester used, 119 mg product obtained (99% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 1.17 (m, 4H), 1.43 (m, 10H), 1.88(dd, J=8.31 5.4 Hz, 1H), 2.29 (m, 2H), 2.57 (dd, J=13.7, 6.9 Hz, 1H),2.95 (m, 1H), 3.87 (m, 2H), 4.42 (dd, J=9.9, 6.9 Hz, 1H), 5.13 (d,J=10.3 Hz, 1H), 5.31 (dd, J=17.1, 1.2 Hz, 1H), 5.78 (m, 2H), 7.35 (d,J=5.9 Hz, 1H), 7.69 (dd, J=8.7, 2.1 Hz, 1H), 7.84 (d, J=8.8 Hz, 1H),7.99 (d, J=5.9 Hz, 1H), 8.12 (d, J=1.7 Hz, 1H); MS: (M+Na)⁺ 627.

Step 5:

Modifications: 115 mg2-(1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-(7-chloro-isoquinolin-1-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester used, 36 mg product obtained (25% yield).

Product:

Data: MS: (M+Na)⁺ 740.

Example 259 Preparation of Compound 259

Compound 259 was prepared by following Scheme 1 of Example 250 exceptthat 4-methylcinnamic acid was used in place of 3-phenyl-but-2-enoicacid in step 1.

Step 1:

Modifications: 25 g 4-methylcinnamic acid used, 15.3 g product obtained(62% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 2.50 (s, 3H), 6.54 (d, J=7.1 Hz,1H), 7.13 (d, J=7.1 Hz, 1H), 7.49 (m, 2H), 8.22 (s, 1H), 11.49 (s, 1H);MS: (M+H)⁺ 160.

Step 2:

Modifications: 15.3 g 7-methyl-2H-isoquinolin-1-one used, 5.15 g productobtained (30% yield).

Product:

Data: ¹H NMR (400 MHz, CDCl₃) δ ppm 2.58 (s, 3H), 7.56 (m, 2H), 7.73 (d,J=8.3 Hz, 1H), 8.09 (s, 1H), 8.20 (d, J=5.6 Hz, 1H); MS: (M+H)⁺ 178.

Step 3:

Modifications: 205 mg 1-chloro-7-methyl-isoquinoline used, 350 mgproduct obtained (89% yield).

Product:

Data: MS: (M+H) 373.

Step 4:

Modifications: 75 mg4-(7-methyl-isoquinolin-1-yloxy)-pyrrolidine-1,2-dicarboxylic acid1-tert-butyl ester used, 107 mg product obtained (95% yield).

Product:

Data: MS: (M+Na)⁺ 607.

Step 5:

Modifications: 107 mg2-(1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-(7-methyl-isoquinolin-1-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester used, 53 mg product obtained (41% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 1.02 (m, 12H), 1.18 (s, 9H), 1.24(m, 1H), 1.45 (dd, J=9.4, 5.5 Hz, 1H), 1.88 (dd, J=8.2, 5.5 Hz, 1H),2.28 (m, 2H), 2.50 (s, 3H), 2.61 (dd, J=13.8, 6.7 Hz, 1H), 3.34 (s, 1H),4.09 (dd, J=11.7, 3.2 Hz, 1H), 4.23 (s, 1H), 4.42 (d, J=12.0 Hz, 1H),4.57 (dd, J=10.0, 7.1 Hz, 1H), 5.12 (dd, J=10.3, 1.5 Hz, 1H), 5.30 (d,J=17.1 Hz, 1H), 5.76 (m, 1H), 5.87 (s, 1H), 7.28 (d, J=5.9 Hz, 1H), 7.55(d, J=8.3 Hz, 1H), 7.71 (d, J=8.3 Hz, 1H), 7.89 (d, J=5.9 Hz, 1H), 7.93(s, 1H); MS: (M+H)⁺ 698.

Example 260 Preparation of Compound 260

Compound 260 was prepared by following Scheme 1 of Example 250 exceptthat 4-methoxycinnamic acid was used in place of 3-phenyl-but-2-enoicacid in step 1.

Step 1:

Modifications: 33 g using 4-methoxycinnamic acid used, 7 g productobtained (33% yield).

Product:

Data: ¹H NMR (500 MHz, CD₃COCD₃) δ ppm 3.90 (s, 3H), 6.49 (d, J=7.0 Hz,1H), 7.10 (d, J=7.3 Hz, 1H), 7.28 (dd, J=8.6, 2.8 Hz, 1H), 7.57 (d,J=8.9 Hz, 1H), 7.71 (d, J=2.8 Hz, 1H).

Step 2:

Modifications: 4 g 7-methoxy-2H-isoquinolin-1-one used, 3 g productobtained (68% yield).

Product:

Data: ¹H NMR (400 MHz, CDCl₃) δ ppm 3.98 (s, 3H), 7.38 (dd, J=8.9, 2.6Hz, 1H), 7.52 (m, 2H), 7.73 (d, J=8.8 Hz, 1H), 8.16 (d, J=5.4 Hz, 1H).

Step 3:

Modifications: 533 mg 1-chloro-7-methoxy-isoquinoline used, 1115 mgproduct obtained (100% yield).

Product:

Step 4:

Modifications: 78 mg4-(7-methoxy-isoquinolin-1-yloxy)-pyrrolidine-1,2-dicarboxylic acid1-tert-butyl ester used, 108 mg product obtained (99% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 1.17 (m, 4H), 1.40 (m, 1H), 1.43 (s,9H), 1.85 (dd, J=8.1, 5.4 Hz, 1H), 2.21 (m, 2H), 2.51 (dd, J=13.7, 6.6Hz, 1H), 2.93 (s, 1H), 3.80 (m, 2H), 3.94 (s, 3H), 4.41 (dd, J=10.0, 6.6Hz, 1H), 4.57 (s, 1H), 5.11 (d, J=11.3 Hz, 1H), 5.29 (d, J=17.1 Hz, 1H),5.77 (m, 2H), 7.01 (d, J=7.8 Hz, 1H), 7.22 (d, J=5.6 Hz, 1H), 7.32 (d,J=8.1 Hz, 1H), 7.58 (t, J=8.0 Hz, 1H), 7.87 (d, J=5.9 Hz, 1H); MS:(M+H)⁺ 601.

Step 5:

Modifications: 100 mg2-(1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-(7-methoxy-isoquinolin-1-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester used, 30 mg product obtained (25% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃SOCD₃) δ ppm 0.90 (m, 2H), 0.95 (s, 9H), 1.05(m, 1H), 1.12 (s, 9H), 1.35 (m, 2H), 1.70 (m, 1H), 2.18 (m, 1H), 2.92(m, 1H), 3.86 (s, 3H), 4.00 (m, 2H), 4.27 (d, J=12.0 Hz, 1H), 4.45 (t,J=8.6 Hz, 1H), 5.09 (d, J=10.8 Hz, 1H), 5.23 (d, J=16.9 Hz, 1H), 5.62(m, 1H), 5.79 (s, 1H), 6.55 (d, J=8.1 Hz, 1H), 7.35 (d, J=6.6 Hz, 1H),7.39 (d, J=2.5 Hz, 1H), 7.43 (dd, J=8.8, 2.2 Hz, 1H), 7.84 (d, J=8.8 Hz,1H), 7.88 (d, J=5.9 Hz, 1H); MS: (M+H)⁺ 714.

Example 261 and 262 Preparation of Compounds 261 and 262

Compounds 261 and 262 were prepared by following Scheme 1 of Example 250except that 4-fluoro-3-methoxycinnamic acid was used in place of3-phenyl-but-2-enoic acid in step 1.

Step 1:

Modifications: 19.6 g 4-fluoro-3-methoxycinnamic acid used, 9.5 gproduct obtained (48% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃COCD₃) δ ppm 4.00 (s, 1H), 6.49 (d, J=7.34 Hz,1H), 7.19 (d, J=7.09 Hz, 1H), 7.29 (d, J=8.07 Hz, 1H), 7.86 (d, J=11.74Hz, 1H).

Step 2:

Modifications: 9 g 7-fluoro-6-methoxy-2H-isoquinolin-1-one used, 7 gproduct obtained (70% yield).

Product:

Data: ¹H NMR (400 MHz, CDCl₃) δ ppm 4.04 (s, 3H), 7.17 (d, J=8.07 Hz,1H), 7.48 (d, J=5.62 Hz, 1H), 7.94 (d, J=11.49 Hz, 1H), 8.20 (d, J=5.62Hz, 1H).

Step 3:

Modifications: 222 mg 1-chloro-7-fluoro-6-methoxy-isoquinoline used, 406mg products obtained.

Products:

Step 4:

Modifications: 400 mg mixture of4-(7-fluoro-6-methoxy-isoquinolin-1-yloxy)-pyrrolidine-1,2-dicarboxylicacid 1-tert-butyl ester and4-(1-chloro-6-methoxy-isoquinolin-7-yloxy)-pyrrolidine-1,2-dicarboxylicacid 1-tert-butyl ester used, 700 mg products obtained.

Product:

Step 5:

Modifications: 700 mg mixture of2-(1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-(7-fluoro-6-methoxy-isoquinolin-1-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester and4-(1-chloro-6-methoxy-isoquinolin-7-yloxy)-2-(1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-pyrrolidine-1-carboxylicacid tert-butyl ester used, 79 mg of compound 261 and 80 mg compound 262obtained.

Product:

Data of compound 261: ¹H NMR (400 MHz, CD₃OD) δ ppm 1.07 (m, 12H), 1.25(m, 10H), 1.44 (m, 1H), 1.88 (dd, J=8.1, 5.6 Hz, 1H), 2.25 (m, 2H), 2.60(dd, J=13.7, 6.9 Hz, 1H), 2.94 (m, 1H), 4.02 (m, 4H), 4.22 (s, 1H), 4.43(d, J=12.2 Hz, 1H), 4.53 (dd, J=10.3, 6.6 Hz, 1H), 5.12 (d, J=10.5 Hz,1H), 5.30 (d, J=16.6 Hz, 1H), 5.75 (m, 1H), 5.84 (s, 1H), 7.28 (d, J=5.9Hz, 1H), 7.37 (d, J=8.1 Hz, 1H), 7.75 (d, J=11.7 Hz, 1H), 7.91 (d, J=5.9Hz, 1H); MS: (M+Na)⁺ 754.

Data of compound 262: ¹H NMR (400 MHz, CD₃OD) δ ppm 1.07 (m, 12H), 1.25(m, 10H), 1.44 (m, J=9.41, 5.26 Hz, 1H), 1.87 (dd, J=8.31, 5.38 Hz, 1H),2.24 (q, J=8.72 Hz, 2H), 2.57 (dd, J=13.82, 7.21 Hz, 1H), 2.94 (m, 1H),3.97 (d, J=5.14 Hz, 3H), 4.09 (m, J=11.00 Hz, 1H), 4.24 (s, 1H), 4.32(m, 1H), 4.50 (m, J=16.87 Hz, 1H), 5.12 (dd, J=10.52, 1.71 Hz, 1H), 5.30(dd, J=17.12, 1.47 Hz, 1H), 5.38 (s, 1H), 5.76 (m, 1H), 7.39 (s, 1H),7.63 (s, 1H), 7.66 (d, J=5.87 Hz, 1H), 8.07 (d, J=5.62 Hz, 1H); MS:(M+H)⁺ 732.

Example 263 Preparation of Compound 263

Compound 263 was prepared by following Scheme 1 of Example 250 exceptstep 1 and step 2.

Step 3:

Modifications: 176 mg 1-chloro-8-methyl-isoquinoline used, 370 mgproduct obtained (100 mg % yield).

Product:

Step 4:

Modifications: 149 mg8-methyl-isoquinolin-1-yloxy)-pyrrolidine-1,2-dicarboxylic acid1-tert-butyl ester used, 230 mg product obtained (99% yield)

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 1.13 (m, 4H), 1.42 (m, 10H), 1.87(dd, J=8.2, 5.3 Hz, 1H), 2.25 (m, 2H), 2.58 (dd, J=13.9, 6.9 Hz, 1H),2.83 (s, 3H), 2.96 (m, 1H), 3.85 (m, 2H), 4.38 (dd, J=10.2, 6.7 Hz, 1H),5.12 (dd, J=10.4, 1.6 Hz, 1H), 5.30 (dd, J=17.1, 1.2 Hz, 1H), 5.76 (m,2H), 7.28 (d, J=5.9 Hz, 1H), 7.36 (d, J=6.9 Hz, 1H), 7.53 (t, J=7.7 Hz,1H), 7.62 (m, 1H), 7.88 (d, J=5.6 Hz, 1H); MS: (M+Na)⁺ 607.

Step 5:

Modifications: 220 mg2-(1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-(8-methyl-isoquinolin-1-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester used, 90 mg product obtained (35% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 1.05 (m, 12H), 1.24 (m, 10H), 1.44(dd, J=9.3, 5.4 Hz, 1H), 1.87 (dd, J=8.1, 5.4 Hz, 1H), 2.25 (m, 2H),2.60 (dd, J=13.9, 7.3 Hz, 1H), 2.77 (s, 3H), 2.94 (m, 1H), 4.04 (dd,J=11.9, 3.1 Hz, 1H), 4.27 (d, J=9.5 Hz, 1H), 4.42 (d, J=12.0 Hz, 1H),4.54 (dd, J=10.8, 7.1 Hz, 1H), 5.12 (d, J=10.3 Hz, 1H), 5.28 (d, J=17.1Hz, 1H), 5.75 (m, 1H), 5.95 (s, 1H), 6.63 (d, J=9.1 Hz, 1H), 7.28 (m,2H), 7.50 (t, J=7.7 Hz, 1H), 7.60 (d, J=7.8 Hz, 1H), 7.89 (d, J=5.6 Hz,1H); MS: (M+Na)⁷²⁰.

Example 264 Preparation of Compound 264

Compound 264 was prepared by following Scheme 1 of Example 250 exceptstep 1 and step 2.

Step 3:

Modifications: 203 mg 1-chloro-8-methoxy-isoquinoline used, 340 mgproduct obtained (85% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃SOCD₃) δ ppm 1.34, 1.36 (2s, 9H, rotamers),2.26 (m, 1H), 2.49 (m, 1H), 3.67 (m, 2H), 3.86 (s, 3H), 4.31 (m, 1H),5.67 (br s, 1H), 7.04 (d, J=7.8 Hz, 1H), 7.30 (d, J=5.9 Hz, 1H), 7.38(d, J=8.1 Hz, 1H), 7.62 (t, J=8.0 Hz, 1H), 7.93 (d, J=5.6 Hz, 1H), 12.64(s, 1H); MS: (M+Na)⁺ 411.

Step 4:

Modifications: 78 mg8-methoxy-isoquinolin-1-yloxy)-pyrrolidine-1,2-dicarboxylic acid1-tert-butyl ester used, 115 mg product obtained (96% yield).

Product:

Step 5:

Modifications: 110 mg2-(1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-(8-methoxy-isoquinolin-1-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester used, 45 mg product obtained (34% yield).

Product:

Data: MS: (M+H)⁺ 714.

Example 265 and 266 Preparation of Compounds 265 and 266

Compounds 265 and 266 were prepared by following Scheme 1 of Example 250except that 3-(2,3-dihydro-benzofuran-7-yl)-acrylic acid was used inplace of 3-phenyl-but-2-enoic acid in step 1.

Step 1:

Modifications: 3.8 g 3-(2,3-dihydro-benzofuran-7-yl)-acrylic acid used,2 g product obtained (53% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 3.37 (t, J=9.05 Hz, 1H), 4.73 (t,J=9.05 Hz, 2H), 6.67 (d, J=7.09 Hz, 1H), 7.10 (d, J=7.09 Hz, 1H), 7.37(d, J=8.07 Hz, 1H), 7.81 (d, J=8.07 Hz, 1H); MS: (M+H)⁺ 188.

Step 2:

Modifications: 1.87 g 2,3-dihydro-7H-furo[2,3-f]isoquinolin-6-one used,1.84 g product obtained (90% yield).

Product:

Data: ¹H NMR (400 Hz, CDCl₃) δ ppm 3.43 (t, J=9.05 Hz, 2H), 4.82 (t,J=9.05 Hz, 2H), 7.52 (d, J=8.56 Hz, 1H), 7.66 (d, J=5.62 Hz, 1H), 7.84(d, J=8.31 Hz, 1H), 8.19 (d, J=5.62 Hz, 1H); MS (M+H)⁺ 206.

Step 3:

Modifications: 206 mg 6-chloro-2,3-dihydro-furo[2,3-f]isoquinoline used,300 mg products mixture obtained.

Products:

Step 4:

Modifications: 240 mg step 3 products mixture used, 350 mg productsmixture obtained.

Products:

Step 5:

Modifications: 331 mg step 4 products mixture used, 240 mg of compound265 and 24 mg of compound 266 obtained.

Products:

Data of compound 265: ¹H NMR (400 Hz, CD₃OD) δ ppm 0.99 (m, 12H), 1.16(m, H), 1.36 (m, 1H), 1.81 (dd, J=8.07, 5.62 Hz, 1H), 2.18 (m, 2H), 2.54(dd, J=13.69, 6.85 Hz, 1H), 2.87 (m, 1H), 3.31 (t, J=9.05 Hz, 2H), 4.01(m, 1H), 4.18 (s, 1H), 4.36 (d, J=11.74 Hz, 1H), 4.46 (dd, J=10.15, 7.21Hz, 1H), 4.70 (m, 2H), 5.05 (d, J=10.27 Hz, 1H), 5.23 (d, J=16.87 Hz,1H), 5.70 (m, 2H), 7.23 (d, J=5.87 Hz, 1H), 7.31 (d, J=8.31 Hz, 1H),7.63 (d, J=8.31 Hz, 1H), 7.82 (d, J=5.87 Hz, 1H); MS (M+H)⁺ 726.

Data of compound 266: ¹H NMR (400 MHz, CD₃OD) δ ppm 1.06 (m, 12H), 1.24(m, 10H), 1.44 (dd, J=10.03, 5.14 Hz, 1H), 1.88 (dd, J=7.83, 5.38 Hz,1H), 2.27 (m, 2H), 2.65 (dd, J=12.96, 6.36 Hz, 1H), 2.94 (m, 1H), 4.08(dd, J=12.35, 3.30 Hz, 1H), 4.25 (s, 1H), 4.54 (m, 2H), 5.12 (d, J=10.27Hz, 1H), 5.29 (d, J=17.12 Hz, 1H), 5.75 (m, 1H), 5.91 (s, 1H), 7.05 (d,J=1.96 Hz, 1H), 7.72 (m, 2H), 8.02 (m, 2H), 8.11 (d, J=5.87 Hz, 1H),9.19 (s, 1H); MS: (M+H)⁺ 724.

Example 267 and 268 Preparation of Compounds 267 and 268

Compounds 267 and 268 were prepared by following Scheme 1 of Example 250except that 3-(2,3-dihydro-benzofuran-4-yl)-acrylic acid was used inplace of 3-phenyl-but-2-enoic acid in step 1.

Step 1:

Modifications: 1.14 g 3-(2,3-dihydro-benzofuran-4-yl)-acrylic acid used,600 mg product obtained (52% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 3.35 (t, J=8.93 Hz, 2H), 4.74 (t,J=8.93 Hz, 2H), 6.49 (d, J=7.09 Hz, 1H), 6.95 (d, J=8.56 Hz, 1H), 7.25(d, J=7.09 Hz, 1H), 8.13 (d, J=8.80 Hz, 1H); MS (M+H)⁺ 188.

Step 2:

Modifications: 560 mg 1,7-dihydro-2H-furo[3,2-f]isoquinolin-6-one used,380 mg product obtained (48% yield).

Product:

Data: ¹H NMR (400 Hz, CDCl₃) δ ppm 3.47 (t, J=9.05 Hz, 2H), 4.84 (t,J=9.05 Hz, 2H), 7.24 (d, J=8.56 Hz, 1H), 7.33 (d, J=5.87 Hz, 1H), 8.20(m, 2H); MS (M+H)⁺ 206.

Step 3:

Modifications: 105 mg 6-chloro-1,2-dihydro-furo[3,2-f]isoquinoline used,390 mg products mixture obtained.

Products:

Step 4:

Modifications: 216 mg step 3 products mixture used, 330 mg productsmixture obtained.

Products:

Step 5:

Modifications: 330 mg step 4 products mixture used, 140 mg of compound267 and mg of compound 268 obtained.

Products:

Data of compound 267: ¹H NMR (400 Hz, CD₃OD) δ ppm 1.07 (m, 12H), 1.24(m, 10H), 1.43 (m, 1H), 1.88 (dd, J=8.07, 5.38 Hz, 1H), 2.26 (m, 2H),2.61 (dd, J=13.69, 7.09 Hz, 1H), 2.94 (m, 1H), 3.42 (t, J=9.05 Hz, 2H),4.05 (dd, J=11.86, 3.55 Hz, 1H), 4.24 (s, 1H), 4.50 (m, 2H), 4.77 (t,J=8.93 Hz, 2H), 5.12 (m, 1H), 5.29 (d, J=17.12 Hz, 1H), 5.76 (m, 2H),7.03 (d, J=8.80 Hz, 1H), 7.12 (d, J=6.11 Hz, 1H), 7.91 (d, J=5.87 Hz,1H), 8.06 (d, J=8.80 Hz, 1H); MS: (M+H)⁺ 726.

Data of compound 268: ¹H NMR (400 Hz, CD₃OD) δ ppm 1.06 (m, 12H), 1.19(s, 9H) 1.26 (m, 1H), 1.44 (m, 1H), 1.88 (dd, J=8.07, 5.62 Hz, 1H), 2.24(d, J=8.56 Hz, 2H), 2.64 (m, 1H), 2.95 (m, 1H), 4.07 (m, J=3.42 Hz, 1H),4.24 (s, 1H), 4.54 (m, 2H), 5.12 (d, J=10.52 Hz, 1H), 5.30 (d, J=17.12Hz, 1H), 5.76 (m, 1H), 5.91 (s, 1H), 7.39 (d, J=1.47 Hz, 1H), 7.68 (m,2H), 7.96 (d, J=1.96 Hz, 1H), 8.12 (m, 2H); MS: (M+H)⁺ 724.

Example 269 Preparation of Compound 269

Step 1:

A solution of 2-trifluormethoxycinnamic acid (11.6 g),diphenylphosphoryl azide (13.75 g), and triethylamine (7.07 g) inbenzene (50 mL) was stirred for 1 h. After filtration through a silicagel plug washing with benzene and concentration, the residue wasdissolved in diphenylmethane (80 mL) and refluxed for 3 h. After coolingto rt, solids were collected through a plug washing with benzene anddried to give 5.1 g (44%) of the desired product as a solid. ¹H NMR (400MHz, CD₃OD) δ ppm 6.79 (d, J=7.3 Hz, 1H), 7.29 (d, J=7.3 Hz, 1H), 7.57(t, J=8.1 Hz, 1H), 7.70 (d, J=7.8 Hz, 1H), 8.30 (d, J=8.1 Hz, 1H); MS:(M+H) 230.

Step 2:

A solution of 5-trifluoromethoxy-2H-isoquinolin-1-one (4.58 g) in POCl₃(50 mL) was refluxed for 3 h. After cooling and concentration, theresidue was based with 5 N NaOH and extracted with CH₂Cl₂. The organiclayer was washed with brine and dried over MgSO₄. After concentration,purification by flash chromatography of Biotage with 5% ethyl acetate inhexanes gave 4.347 g (88%) of the desired product as a solid. ¹H NMR(400 MHz, CDCl₃) δ ppm 7.66 (m, 2H), 7.87 (d, J=5.9 Hz, 1H), 8.31 (m,1H), 8.37 (d, J=5.9 Hz, 1H); MS: (M+H)⁺ 248.

Step 3:

To a suspension of{1-[2-(1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl}-carbamicacid tert-butyl ester (56 mg), 1-chloro-5-trifluoromethoxy-isoquinoline(25 mg), and LaCl₃ (25 mg) in DMF (1 mL) at −78° C. was added tert-BuOK(0.5 mL, 1 M in THF) and warmed to rt. After stirring for 30 min, thereaction was quenched with saturated NH₄Cl solution and extracted withethyl acetate. After concentration, purification by prep HPLC gave 35 mg(46%) of the desired compound 269 as a solid. ¹H NMR (400 MHz, CD₃OD) δppm 1.03 (m, 12H), 1.24 (m, 10H), 1.44 (dd, J=9.7, 5.3 Hz, 1H), 1.88(dd, J=8.1, 5.6 Hz, 1H), 2.28 (m, 2H), 2.64 (dd, J=13.7, 7.1 Hz, 1H),2.94 (m, 1H), 4.09 (m, 1H), 4.21 (d, J=9.3 Hz, 1H), 4.53 (m, 2H), 5.12(d, J=11.5 Hz, 1H), 5.30 (d, J=17.1 Hz, 1H), 5.75 (m, 1H), 5.92 (m, 1H),6.60 (d, J=9.5 Hz, 1H), 7.49 (d, J=6.1 Hz, 1H), 7.60 (m, 1H), 7.69 (d,J=7.3 Hz, 1H), 8.11 (d, J=6.1 Hz, 1H), 8.22 (d, J=8.3 Hz, 1H); MS:(M+Na)⁺ 790.

Example 270 Preparation of Compound 270

Compound 270 was prepared by following Scheme 2 of Example 269 exceptthat 2-trifluoromethylcinnamic acid was used in place of2-trifluormethoxycinnamic acid in step 1.

Step 1:

Modifications: 10 g 2-trifluoromethylcinnamic acid used, 5 g productobtained (50% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 6.83 (m, 1H), 7.33 (d, J=7.58 Hz,1H), 7.63 (t, J=7.83 Hz, 1H), 8.09 (d, J=7.58 Hz, 1H), 8.57 (d, J=8.07Hz, 1H).

Step 2:

Modifications: 4.4 g 5-trifluoromethyl-2H-isoquinolin-1-one used, 3.5 gproduct obtained (73% yield).

Product:

Data: ¹H NMR (400 MHz, CDCl₃) δ ppm 7.75 (t, J=7.95 Hz, 1H), 7.90 (m,1H), 8.12 (d, J=7.34 Hz, 1H), 8.41 (d, J=6.11 Hz, 1H), 8.60 (d, J=8.56Hz, 1H).

Step 3:

Modifications: 46 mg 1-chloro-5-trifluoromethyl-isoquinoline and 111 mg{1-[2-(1-Cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl}-carbamicacid tert-butyl ester used, 70 mg product obtained (47% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 1.06 (m, 12H), 1.23 (m, 10H), 1.44(dd, J=9.54, 5.38 Hz, 1H), 1.88 (dd, J=8.07, 5.38 Hz, 1H), 2.28 (m, 2H),2.65 (dd, J=13.82, 6.97 Hz, 1H), 2.94 (m, 1H), 4.07 (m, 1H), 4.20 (m,1H), 4.56 (m, 2H), 5.12 (m, 1H), 5.30 (d, J=17.12 Hz, 1H), 5.75 (m, 1H),5.90 (s, 1H), 6.59 (d, J=9.05 Hz, 1H), 7.53 (d, J=4.40 Hz, 1H), 7.65 (t,J=7.83 Hz, 1H), 8.12 (d, J=7.09 Hz, 1H), 8.15 (d, J=6.36 Hz, 1H), 8.50(d, J=8.31 Hz, 1H); MS: (M+Na)⁺ 774.

Example 271 Preparation of Compound 271

Compound 271 was prepared by following Scheme 2 of Example 269 exceptthat 2-chlorocinnamic acid was used in place of2-trifluormethoxycinnamic acid in step 1.

Step 1:

Modifications: 7 g 2-chlorocinnamic acid used, 5 g product obtained (71%yield).

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 3.02 (m, 4H), 3.91 (m, 4H), 6.97 (d,J=7.34 Hz, 1H), 7.18 (d, J=7.34 Hz, 1H), 7.44 (m, 2H), 8.02 (d, J=7.83Hz, 1H); MS (M+H)⁺ 231.

Step 2:

Modifications: 2.2 g 5-morpholin-4-yl-2H-isoquinolin-1-one used, 2.1 gproduct obtained (87% yield).

Product:

Data: ¹H NMR (400 MHz, CCl₃D) δ ppm 3.09 (m, 4H), 3.97 (m, 4H), 7.32 (d,J=7.58 Hz, 1H), 7.60 (m, 1H), 7.91 (d, J=5.87 Hz, 1H), 8.06 (d, J=8.56Hz, 1H), 8.26 (d, J=5.87 Hz, 1H).

Step 3:

Modifications: 50 mg 1-chloro-5-morpholin-4-yl-isoquinoline and 111 mg{1-[2-(1-Cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl}-carbamicacid tert-butyl ester used, 40 mg product obtained (26% yield).

Product:

Data: ¹H NMR (500 MHz, CD₃OD) δ ppm 1.07 (m, 12H), 1.26 (m, 10H), 1.44(d, J=7.93 Hz, 1H), 1.88 (dd, J=7.93, 5.19 Hz, 1H), 2.25 (m, 2H), 2.62(dd, J=13.73, 7.02 Hz, 1H), 2.94 (m, 1H), 3.06 (d, J=3.97 Hz, 4H), 3.94(m, 4H), 4.07 (d, J=14.04 Hz, 1H), 4.25 (s, 1H), 4.45 (d, J=12.21 Hz,1H), 4.52 (m, 1H), 5.12 (d, J=9.46 Hz, 1H), 5.29 (d, J=16.79 Hz, 1H),5.75 (m, 1H), 5.85 (s, 1H), 7.34 (d, J=7.32 Hz, 1H), 7.45 (t, J=7.78 Hz,1H), 7.59 (d, J=6.10 Hz, 1H), 7.91 (d, J=7.63 Hz, 1H), 7.97 (d, J=5.80Hz, 1H).

Example 272 Preparation of Compound 272

Compound 272 was prepared by following Scheme 2 of Example 269 exceptthat 2,3-dimethoxycinnamic acid was used in place of2-trifluormethoxycinnamic acid in step 1.

Step 1:

Modifications: 10.4 g 2,3-dimethoxycinnamic acid used, 4.1 g productobtained (40% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 3.86 (s, 3H), 3.96 (s, 3H), 6.82 (d,J=7.2 Hz, 1H), 7.10 (d, J=7.2 Hz, 1H), 7.28 (d, J=8.8 Hz, 1H), 8.07 (d,J=8.8 Hz, 1H);

MS: (M+H)⁺ 206.

Step 2:

Modifications: 4.1 g 5,6-dimethoxy-2H-isoquinolin-1-one used, 4.03 gproduct obtained (90% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 3.97 (s, 3H), 4.05 (s, 3H), 7.65 (d,J=9.29 Hz, 1H), 7.90 (dd, J=5.87, 0.98 Hz, 1H), 8.12 (m, 2H).

Step 3:

Modifications: 22 mg 1-chloro-5,6-dimethoxy-isoquinoline and 56 mg{1-[2-(1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl}-carbamicacid tert-butyl ester used, 31 mg product obtained (42% yield).

Product:

Data: ¹H NMR (500 MHz, CD₃OD) δ ppm 1.06 (m, 12H), 1.26 (m, 10H), 1.44(s, 1H), 1.88 (d, J=7.32 Hz, 1H), 2.24 (s, 2H), 2.60 (m, 1H), 2.94 (m,1H), 3.92 (s, 3H), 3.99 (s, 3H), 4.06 (d, J=11.90 Hz, 1H), 4.23 (s, 1H),4.43 (d, J=10.68 Hz, 1H), 4.53 (m, 1H), 5.12 (d, J=10.38 Hz, 1H), 5.30(d, J=17.40 Hz, 1H), 5.77 (m, 2H), 7.35 (d, J=9.16 Hz, 1H), 7.46 (d,J=5.80 Hz, 1H), 7.89 (d, J=5.80 Hz, 1H), 7.97 (d, J=8.85 Hz, 1H).

Example 273 Preparation of Compound 273

Compound 273 was prepared by following Scheme 2 of Example 269 exceptthat 4-chloro-3-methoxycinnamic acid was used in place of2-trifluormethoxycinnamic acid in step 1.

Step 1:

Modifications: 2.5 g 4-chloro-3-methoxycinnamic acid used, 1.2 g productobtained (48% yield).

Product:

Data: ¹HNMR (400 MHz, CD₃OD) δ 4.00 (s, 3H), 6.64 (d, J=7.09 Hz, 1H),7.15 (d, J=7.34 Hz, 1H), 7.21 (s, 1H), 8.22 (s, 1H).

Step 2:

Modifications: 1.05 g 7-Chloro-6-methoxy-2H-isoquinolin-1-one used, 0.8g product obtained (70% yield).

Product:

Data: ¹H NMR (400 Hz, CDCl₃) δ ppm 4.05 (s, 3H), 7.13 (s, 1H), 7.48 (d,J=5.38 Hz, 1H), 8.21 (d, J=5.62 Hz, 1H), 8.34 (s, 1H); MS: (M+H)⁺ 229.

Step 3:

Modifications: 44 mg 1,7-dichloro-6-methoxy-isoquinoline and 113 mg{1-[2-(1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl}-carbamicacid tert-butyl ester used, 25 mg product obtained (17% yield)

Product:

Data: ¹H NMR (400 Hz, CD₃OD) δ ppm 1.07 (m, 12H), 1.24 (m, 10H), 1.44(dd, J=9.54, 5.38 Hz, 1H), 1.88 (dd, J=8.07, 5.38 Hz, 1H), 2.26 (m, 1H),2.60 (m, J=13.69, 6.85 Hz, 1H), 2.94 (m, 2H), 3.98 (s, 3H), 4.06 (m,1H), 4.20 (m, 1H), 4.42 (d, J=12.23 Hz, 1H), 4.57 (m, 1H), 5.12 (d,J=11.74 Hz, 1H), 5.30 (d, J=17.36 Hz, 1H), 5.76 (m, 1H), 5.86 (s, 1H),7.28 (d, J=5.62 Hz, 1H), 7.33 (s, 1H), 7.92 (d, J=5.87 Hz, 1H), 8.09 (s,1H); MS: (M+H)⁺ 749.

Example 274 Preparation of Compound 274

Compound 274 was prepared by following Scheme 2 of Example 269 exceptthat 2-fluoro-3-cinnamic acid was used in place of2-trifluormethoxycinnamic acid in step 1.

Step 1:

Modifications: 3.92 g 2-fluoro-3-cinnamic acid used, 2.4 g productobtained (61% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 4.00 (s, 3H), 6.72 (m, 1H), 7.16 (d,J=7.34 Hz, 1H), 7.35 (t, J=8.44 Hz, 1H), 8.09 (d, J=8.80 Hz, 1H).

Step 2:

Modifications: 1.93 g 5-fluoro-6-methoxy-2H-isoquinolin-1-one used,1.688 g product obtained (80% yield).

Product:

Data: ¹H NMR (CDCl₃) δ ppm 4.08 (s, 3H), 7.44 (dd, J=9.29, 7.83 Hz, 1H),7.75 (d, J=5.87 Hz, 1H), 8.12 (d, J=9.29 Hz, 1H), 8.22 (d, J=5.87 Hz,1H); MS: (M+H)⁺ 212.

Step 3:

Modifications: 41 mg 1-chloro-5-fluoro-6-methoxy-isoquinoline and 133 mg{1-[2-(1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl}-carbamicacid tert-butyl ester used, 70 mg product obtained (48% yield).

Product:

Data: ¹H NMR (CD₃OD) δ ppm 1.06 (m, 13H), 1.21 (s, 9H), 1.44 (dd,J=9.78, 5.38 Hz, 1H), 1.88 (dd, J=8.19, 5.50 Hz, 1H), 2.24 (d, J=9.29Hz, 2H), 2.62 (d, J=13.94 Hz, 1H), 2.94 (m, 1H), 4.05 (m, 4H), 4.22 (d,J=9.29 Hz, 1H), 4.45 (m, 1H), 4.54 (dd, J=9.66, 7.21 Hz, 1H), 5.12 (d,J=10.52 Hz, 1H), 5.30 (d, J=16.87 Hz, 1H), 5.76 (m, 1H), 5.86 (s, 1H),7.39 (m, 2H), 7.95 (d, J=6.11 Hz, 1H), 8.00 (d, J=9.29 Hz, 1H).

Example 275 Preparation of Compound 275

Compound 2 was prepared by following Scheme 2 of Example 269 except that2-chloro-3-methoxycinnamic acid was used in place of2-trifluormethoxycinnamic acid in step 1.

Step 1:

Modifications: 658 mg 2-chloro-3-methoxycinnamic acid used, 360 mgproduct obtained (54% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 4.02 (s, 3H), 6.91 (d, J=7.34 Hz,1H), 7.23 (d, J=7.58 Hz, 1H), 7.35 (d, J=9.05 Hz, 1H), 8.27 (d, J=9.05Hz, 1H).

Step 2:

Modifications: 350 mg 5-chloro-6-methoxy-2H-isoquinolin-1-one used, 300mg product obtained (80% yield).

Product:

Data: ¹H NMR (400 Hz, CDCl₃) δ ppm 4.09 (s, 3H), 7.43 (d, J=9.29 Hz,1H), 7.93 (d, J=6.11 Hz, 1H), 8.30 (m, 2H); MS (M+H)⁺ 229.

Step 3:

Modifications: 68 mg 1,5-dichloro-6-methoxy-isoquinoline and 167 mg{1-[2-(1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl}-carbamicacid tert-butyl ester used, 130 mg product obtained (60% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 1.06 (m, 12H), 1.25 (m, 10H), 1.46(d, J=5.62 Hz, 1H), 1.88 (dd, J=8.07, 5.62 Hz, 1H), 2.27 (m, 2H), 2.62(m, 1H), 2.94 (m, 1H), 4.05 (m, 4H), 4.22 (d, J=9.05 Hz, 1H), 4.46 (d,J=11.49 Hz, 1H), 4.54 (dd, J=9.78, 6.36 Hz, 1H), 5.13 (d, J=10.52 Hz,1H), 5.30 (d, J=15.89 Hz, 1H), 5.76 (m, 1H), 5.86 (s, 1H), 7.40 (d,J=9.29 Hz, 1H), 7.55 (d, J=6.36 Hz, 1H), 8.01 (d, J=6.36 Hz, 1H), 8.20(d, J=9.29 Hz, 1H); MS: (M+H)⁺ 749.

Example 276 Preparation of Compound 276

Compound 276 was prepared by following Scheme 2 of Example 269 exceptthat 3-chloro-2-methoxycinnamic acid was used in place of2-trifluormethoxycinnamic acid in step 1.

Step 1:

Modifications: 4.24 g 3-chloro-2-methoxycinnamic acid used, 2.4 gproduct obtained (57% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 3.93 (s, 1H), 6.85 (d, J=7.34 Hz,1H), 7.24 (d, J=7.34 Hz, 1H), 7.52 (d, J=8.80 Hz, 1H), 8.03 (d, J=8.80Hz, 1H); MS: (M+H)⁺ 210.

Step 2:

Modifications: 2.09 g 6-chloro-5-methoxy-2H-isoquinolin-1-one used, 1.9g product obtained (83% yield).

Product:

Data: ¹H NMR (400 Hz, CDCl₃) δ ppm 4.03 (s, 2H), 7.63 (d, J=9.05 Hz,1H), 7.86 (d, J=5.14 Hz, 1H), 8.06 (d, J=9.05 Hz, 1H), 8.32 (d, J=5.62Hz, 1H); MS: (M+H)⁺ 229.

Step 3:

Modifications: 91 mg 1,6-dichloro-5-methoxy-isoquinoline and 226 mg{1-[2-(1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl}-carbamicacid tert-butyl ester used, 114 mg product obtained (38% yield).

Product:

Data: ¹H NMR (400 Hz, CD₃OD) δ ppm 1.06 (m, 12H), 1.23 (m, 10H), 1.44(t, J=6.72 Hz, 1H), 1.88 (dd, J=7.95, 5.26 Hz, 1H), 2.25 (m, 2H), 2.62(dd, J=13.33, 6.48 Hz, 1H), 2.94 (m, 1H), 3.98 (s, 3H), 4.03 (m, 1H),4.20 (m, 1H), 4.51 (m, 2H), 5.12 (d, J=10.52 Hz, 1H), 5.32 (s, 1H), 5.75(m, 1H), 5.87 (s, 1H), 7.50 (m, 2H), 7.95 (d, J=8.80 Hz, 1H), 8.06 (d,J=5.87 Hz, 1H); MS (MH⁺) 749.

Example 277 Preparation of Compound 277

Compound 277 was prepared by following Scheme 2 of Example 269 exceptthat 3-(4-chloro-phenyl)-3-methoxy-acrylic acid was used in place of2-trifluormethoxycinnamic acid in step 1.

Step 1:

Modifications: 4.24 g 3-(4-chloro-phenyl)-3-methoxy-acrylic acid used,130 mg product obtained (3% yield)

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 3.96 (s, 3H), 7.19 (dd, J=8.80, 2.45Hz, 1H), 7.28 (d, J=2.45 Hz, 1H), 7.34 (s, 1H), 8.25 (d, J=9.05 Hz, 1H);MS: (M+H)⁺ 210.

Step 2:

Modifications: 105 mg 7-chloro-4-methoxy-2H-isoquinolin-1-one used, 60mg product obtained (71% yield).

Product:

Data: ¹H NMR (400 Hz, CDCl₃) δ ppm 4.05 (s, 3H), 7.67 (dd, J=8.80, 1.96Hz, 1H), 7.80 (s, 1H), 8.16 (d, J=9.05 Hz, 1H), 8.24 (d, J=1.96 Hz, 1H);MS: (M+H)⁺ 229.

Step 3:

Modifications: 46 mg 1,7-dichloro-4-methoxy-isoquinoline and 113 mg{1-[2-(1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl}-carbamicacid tert-butyl ester used, 50 mg product obtained (31% yield).

Product:

Data: ¹H NMR (400 Hz, CD₃OD) δ ppm 1.06 (m, 11H), 1.16 (s, 9H), 1.24 (m,2H), 1.44 (dd, J=9.54, 5.38 Hz, 1H), 1.88 (dd, J=8.07, 5.62 Hz, 1H),2.28 (m, 2H), 2.59 (dd, J=13.69, 6.85 Hz, 1H), 2.94 (m, 1H), 4.00 (s,3H), 4.05 (d, J=11.74 Hz, 1H), 4.19 (s, 1H), 4.43 (d, J=11.49 Hz, 1H),4.56 (dd, J=10.03, 6.85 Hz, 1H), 5.12 (d, J=11.49 Hz, 1H), 5.30 (d,J=17.12 Hz, 1H), 5.76 (m, 2H), 7.57 (s, 1H), 7.67 (d, J=8.56 Hz, 1H),8.04 (s, 1H), 8.08 (d, J=8.80 Hz, 1H); MS: (M+H)⁺ 749.

Example 278 Preparation of Compound 278

Compound 278 was prepared by following Scheme 2 of Example 269 exceptstep 1.

Step 1:

Modifications: A mixture of 6-methoxy-2H-isoquinolin-1-one (700 mg) andNCS (532 mg) in MeCN (10 mL) was refluxed for 3 h. Filtration gave 600mg (72%) of the desired product as a solid.

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 3.96 (s, 1H), 7.19 (dd, J=8.80, 2.45Hz, 1H), 7.28 (d, J=2.45 Hz, 1H), 7.34 (s, 1H), 8.25 (d, J=9.05 Hz, 1H);MS: (M+H)⁺ 210.

Step 2:

Modifications: 500 mg 4-chloro-6-methoxy-2H-isoquinolin-1-one used, 400mg product obtained.

Product:

Data: ¹H NMR (400 Hz, CDCl₃) δ ppm 4.01 (s, 3H), 7.35 (d, J=2.45 Hz,1H), 7.41 (d, J=2.45 Hz, 1H), 8.24 (d, J=9.29 Hz, 1H), 8.27 (s, 1H); MS:(M+H)⁺ 229.

Step 3:

Modifications: 42 mg 1,4-dichloro6-methoxy-isoquinoline and 117 mg{1-[2-(1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl}-carbamicacid tert-butyl ester used, 70 mg product obtained (47% yield).

Product:

Data: ¹H NMR (400 Hz, CD₃OD) δ ppm 1.05 (m, 12H), 1.25 (m, 10H), 1.44(m, 1H), 1.88 (dd, J=8.07, 5.62 Hz, 1H), 2.24 (m, 2H), 2.61 (dd,J=13.82, 6.72 Hz, 1H), 2.94 (m, 1H), 3.97 (s, 3H), 4.04 (dd, J=11.74,2.69 Hz, 1H), 4.21 (s, 1H), 4.49 (m, 2H), 5.12 (d, J=10.52 Hz, 1H), 5.29(d, J=17.12 Hz, 1H), 5.75 (m, 2H), 7.19 (d, J=8.80 Hz, 1H), 7.37 (s,1H), 8.00 (s, 1H), 8.13 (d, J=9.05 Hz, 1H); MS: (M+H)⁺ 749.

Example 279 Preparation of Compound 279

Compound 279 was prepared by following Scheme 2 of Example 269 exceptthat 3-methoxy-3-(3-methoxy-phenyl)-acrylic acid was used in place of2-trifluormethoxycinnamic acid in step 1.

Step 1:

Modifications: 4.24 g 3-methoxy-3-(3-methoxy-phenyl)-acrylic acid used,400 mg product obtained (10% yield).

Product:

Step 2:

Modifications: 400 mg 4,6-dimethoxy-2H-isoquinolin-1-one used, 300 mgproduct obtained (69% yield).

Product:

Data: ¹H NMR (400 Hz, CDCl₃) δ ppm 3.97 (s, 3H), 4.05 (s, 3H), 7.31 (dd,J=9.17, 2.57 Hz, 1H), 7.45 (d, J=2.69 Hz, 1H), 7.75 (s, 1H), 8.16 (d,J=9.29 Hz, 1H); MS: (M+H)⁺ 224.

Step 3:

Modifications: 89 mg 1-chloro-4,6-dimethoxy-isoquinoline and 223 mg{1-[2-(1-Cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl}-carbamicacid tert-butyl ester used, 160 mg product obtained (54% yield).

Product:

Data: ¹H NMR (400 Hz, CD₃OD) δ ppm 1.07 (m, 12H), 1.21 (m, 10H), 1.43(m, 1H), 1.87 (dd, J=8.07, 5.62 Hz, 1H), 2.24 (m, 2H), 2.58 (dd,J=13.57, 6.97 Hz, 1H), 2.94 (m, 1H), 3.92 (s, 3H), 3.99 (s, 3H), 4.04(dd, J=11.74, 2.93 Hz, 1H), 4.24 (s, 1H), 4.39 (d, J=11.98 Hz, 1H), 4.50(m, 1H), 5.12 (d, J=10.52 Hz, 1H), 5.29 (d, J=16.87 Hz, 1H), 5.75 (m,2H), 7.12 (d, J=9.05 Hz, 1H), 7.40 (d, J=2.20 Hz, 1H), 7.48 (s, 1H),8.04 (d, J=9.05 Hz, 1H); MS: (M+H)⁺ 744.

Example 280 Preparation of Compound 280

Compound 280 was prepared by following Scheme 2 of Example 269 exceptthat 3-(3-difluoromethoxy-phenyl)-acrylic acid was used in place of2-trifluormethoxycinnamic acid in step 1.

Step 1:

Modifications: 4.28 g 3-(3-difluoromethoxy-phenyl)-acrylic acid used,3.1 g product obtained (72% yield).

Product:

Data: MS: (M+H)⁺ 212.

Step 2:

Modifications: 2 g 6-difluoromethoxy-2H-isoquinolin-1-one used, 1.5 gproduct obtained (61% yield).

Product:

Data: ¹H NMR (400 Hz, CDCl₃) δ ppm 6.69 (t, J=72.75 Hz, 1H), 7.49 (m,2H), 8.28 (d, J=5.62 Hz, 1H), 8.36 (d, J=9.05 Hz, 1H); MS: (M+H)⁺ 230.

Step 3:

Modifications: 46 mg 1-chloro-6-difluoromethoxy-isoquinoline and 113 mg{1-[2-(1-Cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl}-carbamicacid tert-butyl ester used, 8 mg product obtained (5% yield).

Product:

Data: ¹H NMR (400 Hz, CD₃OD) δ ppm 1.05 (m, 12H), 1.23 (m, 10H), 1.44(m, 2H), 1.88 (dd, J=8.19, 5.50 Hz, 1H), 2.30 (m, 2H), 2.67 (d, J=13.94Hz, 1H), 2.93 (m, 1H), 4.07 (d, J=10.27 Hz, 1H), 4.21 (s, 1H), 4.53 (d,J=6.85 Hz, 2H), 5.13 (m, 1H), 5.31 (s, 1H), 5.76 (d, J=47.93 Hz, 2H),7.11 (m, 2H), 7.26 (d, J=6.11 Hz, 1H), 7.81 (d, J=6.11 Hz, 1H), 8.16 (m,1H); MS: (M+H)⁺ 700.

Example 281 Preparation of Compound 281

Compound 281 was prepared by following Scheme 2 of Example 269 exceptthat 3-chloro-3-phenyl-acrylic acid was used in place of2-trifluormethoxycinnamic acid in step 1.

Step 1:

Modifications: 11 g 3-chloro-3-phenyl-acrylic acid used, 3.1 g productobtained (29% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 7.34 (s, 1H), 7.52 (t, J=7.58 Hz,1H), 7.77 (t, J=7.46 Hz, 1H), 7.90 (d, J=8.07 Hz, 1H), 8.39 (d, J=8.07Hz, 1H), 11.37 (s, 1H); MS: MS: (M+H)⁺ 180.

Step 2:

Modifications: 3.1 g 4-chloro-2H-isoquinolin-1-one used, 2.3 g productobtained (66% yield)

Product:

Data: ¹H NMR (400 MHz, CDCl₃) δ ppm 7.77 (ddd, J=8.31, 7.09, 1.22 Hz,1H), 7.88 (ddd, J=8.31, 7.09, 1.22 Hz, 1H), 8.23 (d, J=8.31 Hz, 1H),8.34 (s, 1H), 8.36 (d, J=8.56 Hz, 1H); MS: (M+H)⁺ 198.

Step 3:

Modifications: 20 mg 1,4-dichloro-isoquinoline and 56 mg{1-[2-(1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl}-carbamicacid tert-butyl ester used, 33 mg product obtained (30% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 1.06 (m, 12H), 1.24 (m, 10H), 1.44(dd, J=9.41, 5.26 Hz, 1H), 1.88 (dd, J=7.83, 5.62 Hz, 1H), 2.27 (m, 2H),2.63 (dd, J=13.82, 6.97 Hz, 1H), 2.94 (m, 1H), 4.06 (dd, J=11.49, 2.45Hz, 1H), 4.22 (d, J=9.29 Hz, 1H), 4.53 (m, 2H), 5.12 (d, J=10.76 Hz,1H), 5.29 (d, J=17.12 Hz, 1H), 5.75 (m, 1H), 5.85 (s, 1H), 6.60 (d,J=8.80 Hz, 1H), 7.63 (t, J=7.58 Hz, 1H), 7.86 (t, J=7.70 Hz, 1H), 8.06(s, 1H), 8.11 (d, J=8.56 Hz, 1H), 8.25 (d, J=8.31 Hz, 1H); MS: (M+H)⁺718.

Example 282 Preparation of Compound 282

Compound 2 was prepared by following Scheme 2 of Example 269 except that3-chloro-3-phenyl-acrylic acid was used in place of2-trifluormethoxycinnamic acid in step 1.

Step 1:

Modifications: 20 g 3-chloro-3-phenyl-acrylic acid used, 2 g productobtained (8% yield).

Product:

Data: MS: (M+H)⁺ 230.

Step 2:

Modifications: 2 g 6-trifluoromethoxy-2H-isoquinolin-1-one used, 0.7product obtained (33% yield).

Product:

Data: ¹H NMR (400 MHz, CDCl₃) δ ppm 7.51 (d, J=9.29 Hz, 1H), 7.59 (d,J=5.62 Hz, 1H), 7.64 (s, 1H), 8.31 (d, J=5.62 Hz, 1H), 8.40 (d, J=9.05Hz, 1H); MS: (M+H)⁺ 248.

Step 3:

Modifications: 50 mg 1-chloro-6-trifluoromethoxy-isoquinoline and 113 mg{1-[2-(1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl}-carbamicacid tert-butyl ester used, 42 mg product obtained (27% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 1.05 (m, 12H), 1.24 (m, 10H), 1.44(dd, J=9.17, 5.50 Hz, 1H), 1.88 (dd, J=8.07, 5.62 Hz, 1H), 2.28 (m, 2H),2.63 (dd, J=13.45, 7.09 Hz, 1H), 2.94 (m, 1H), 4.06 (dd, J=11.25, 2.45Hz, 1H), 4.21 (s, 1H), 4.53 (m, 2H), 5.13 (d, J=10.52 Hz, 1H), 5.30 (d,J=17.12 Hz, 1H), 5.75 (m, 1H), 5.89 (s, 1H), 7.39 (m, 2H), 7.72 (s, 1H),8.05 (d, J=5.87 Hz, 1H), 8.31 (d, J=9.05 Hz, 1H), 9.18 (s, 1H); MS:(M+H)⁺ 768.

Example 283 Preparation of Compound 283

Compound 283 was prepared by following Scheme 2 of Example 269 exceptthat 3-(4-fluoro-phenyl)-3-methoxy-acrylic acid was used in place of2-trifluormethoxycinnamic acid in step 1.

Step 1:

Modifications: 3.82 g 3-(4-Fluoro-phenyl)-3-methoxy-acrylic acid used,198 mg product obtained (5% yield).

Product:

Data: MS: (M+H)⁺ 194.

Step 2:

Modifications: 193 mg 7-fluoro-4-methoxy-2H-isoquinolin-1-one used, 199mg product obtained (94% yield).

Product:

Data: ¹H NMR (400 MHz, CDCl₃) δ ppm 4.05 (s, 3H), 7.49 (m, 1H), 7.78 (s,1H), 7.86 (dd, J=9.66, 2.57 Hz, 1H), 8.23 (dd, J=9.29, 5.38 Hz, 1H); MS:(M+H)⁺ 212.

Step 3:

Modifications: 42 mg 1-chloro-7-fluoro-4-methoxy-isoquinoline and 112 mg{1-[2-(1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl}-carbamicacid tert-butyl ester used, 40 mg product obtained (14% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 1.06 (m, 12H), 1.24 (m, 10H), 1.42(m, 1H), 1.87 (dd, J=7.95, 5.50 Hz, 1H), 2.23 (m, 2H), 2.55 (dd,J=13.08, 6.48 Hz, 1H), 2.93 (m, 1H), 4.06 (s, 3H), 4.09 (m, 1H), 4.23(s, 1H), 4.30 (d, J=11.49 Hz, 1H), 4.46 (m, 1H), 5.12 (d, J=10.27 Hz,1H), 5.29 (d, J=17.36 Hz, 1H), 5.40 (s, 1H), 5.76 (m, 1H), 7.46 (d,J=9.05 Hz, 1H), 7.56 (d, J=2.20 Hz, 1H), 7.75 (s, 1H), 8.18 (d, J=9.05Hz, 1H); MS: (M+H)⁺ 749.

Example 284 Preparation of Compound 284

Compound 284 was prepared by following Scheme 2 of Example 269 exceptstep 1.

Step 1:

Modifications: A mixture of 7-methoxy-2H-isoquinolin-1-one (876 mg) andNCS (665 mg) in MeCN (10 mL) was refluxed for 3 h. Filtration gave 500mg (47%) of the desired product as a solid.

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 4.00 (s, 3H), 7.58 (m, 2H), 8.14 (d,J=10.03 Hz, 1H), 8.17 (s, 1H).

Step 2:

Modifications: 418 mg 4-chloro-7-methoxy-2H-isoquinolin-1-one used, 410mg product obtained (90% yield).

Product:

Data: ¹H NMR (400 Hz, CDCl₃) δ ppm 4.00 (s, 3H), 7.49 (dd, J=9.16, 2.44Hz, 1H), 7.55 (d, J=2.44 Hz, 1H), 8.12 (d, J=9.16 Hz, 1H), 8.21 (s, 1H);MS: (M+H)⁺ 229.

Step 3:

Modifications: 42 mg 1,4-dichloro-7-methoxy-isoquinoline and 117 mg{1-[2-(1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl}-carbamicacid tert-butyl ester used, 50 mg product obtained (33% yield).

Product:

Data: ¹H NMR (400 Hz, CD₃OD) δ ppm 1.05 (m, 20H), 1.24 (m, 2H), 1.44 (m,1H), 1.89 (dd, J=8.19, 5.50 Hz, 1H), 2.28 (m, 2H), 2.62 (dd, J=13.69,6.85 Hz, 1H), 2.94 (m, 1H), 3.92 (s, 3H), 4.07 (dd, J=11.98, 3.42 Hz,1H), 4.19 (m, 1H), 4.44 (d, J=11.74 Hz, 1H), 4.58 (dd, J=10.27, 7.09 Hz,1H), 5.12 (m, 1H), 5.31 (d, J=17.12 Hz, 1H), 5.78 (m, 2H), 7.49 (m, 2H),7.91 (s, 1H), 8.02 (m, 1H); MS: (M+H)⁺ 749.

Example 285 Preparation of Compound 285

Compound 285 was prepared by following Scheme 2 of Example 269 exceptthat was used in place of 2-difluormethoxycinnamic acid in step 1.

Step 1 and Step 2:

See compound 256

Step 3:

Modifications: 46 mg 1-chloro-5-difluoromethoxy-isoquinoline and 111 mg{1-[2-(1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl}-carbamicacid tert-butyl ester used, 40 mg product obtained (27% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 1.06 (m, 12H), 1.25 (m, 10H), 1.44(m, 1H, 1.89 (m, 1H), 2.22 (m, 2H), 2.62 (m, 1H), 2.94 (m, 1H), 4.08 (m,1H), 4.23 (d, J=9.54 Hz, 1H), 4.52 (m, 2H), 5.12 (d, J=10.76 Hz, 1H),5.29 (d, J=17.61 Hz, 1H), 5.75 (m, J=10.03 Hz, 1H), 5.88 (s, 1H), 6.60(s, 1H), 7.02 (t, J=73.48 Hz, 1H), 7.52 (m, 3H), 8.07 (m, J=5.75, 5.75Hz, 2H); MS: (M+Na)⁺ 772.

Example 286 Preparation of Compound 286

Compound 286 was prepared by following Scheme 2 of Example 269 exceptthat 3-(2,3-dihydro-benzo[1,4]dioxin-5-yl)-acrylic acid was used inplace of 2-trifluormethoxycinnamic acid in step 1.

Step 1:

Modifications: 4.12 g 3-(2,3-dihydro-benzo[1,4]dioxin-5-yl)-acrylic acidused, 2.2 g product obtained (53% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃OD) δ ppm 4.37 (m, 4H), 6.83 (d, J=7.09 Hz,1H), 7.02 (d, J=8.80 Hz, 1H), 7.12 (d, J=7.34 Hz, 1H), 7.79 (d, J=8.80Hz, 1H); MS: (M+H)⁺ 204.

Step 2:

Modifications: 2.05 g 2,3-dihydro-7H-1,4-dioxa-7-aza-phenanthren-8-oneused, 1.5 g product obtained (68% yield).

Product:

Data: ¹H NMR (400 Hz, CDCl₃) δ ppm 4.42 (m, 4H), 7.24 (d, J=9.05 Hz,1H), 7.77 (d, J=5.87 Hz, 1H), 7.84 (d, J=9.05 Hz, 1H), 8.18 (d, J=5.87Hz, 1H); MS: (M+H)⁺ 222.

Step 3:

Modifications: 88 mg 8-Chloro-2,3-dihydro-1,4-dioxa-7-aza-phenanthreneand 223 mg{1-[2-(1-Cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl}-carbamicacid tert-butyl ester used, 140 mg product obtained (47% yield).

Product:

Data: ¹H NMR (400 Hz, CD₃OD) δ ppm 1.06 (m, 12H), 1.24 (m, 10H), 1.43(dd, J=9.05, 5.14 Hz, 1H), 1.87 (m, 1H), 2.22 (d, J=9.29 Hz, 2H), 2.60(dd, J=13.45, 7.09 Hz, 1H), 2.94 (m, 1H), 4.05 (dd, J=11.62, 2.81 Hz,1H), 4.24 (s, 1H), 4.44 (m, 6H), 5.13 (d, J=17.36 Hz, 1H), 5.29 (d,J=17.36 Hz, 1H), 5.75 (m, 2H), 7.04 (d, J=9.29 Hz, 1H), 7.44 (d, J=5.87Hz, 1H), 7.69 (d, J=9.05 Hz, 1H), 7.88 (d, J=6.11 Hz, 1H); MS: (M+H)⁺742.

Example 287 Preparation of Compound 287

Compound 287 was prepared by following Scheme 2 of Example 269 exceptthat 3-(2,2-difluoro-benzo[1,3]dioxol-4-yl)-acrylic acid was used inplace of 2-trifluormethoxycinnamic acid in step 1.

Step 1:

Modifications: 4.56 g 3-(2,2-difluoro-benzo[1,3]dioxol-4-yl)-acrylicacid used, 2.2 g product obtained (55% yield).

Product:

Data: ¹H NMR (400 MHz, CD₃DOD) δ ppm 6.63 (d, J=7.09 Hz, 1H), 7.29 (d,J=7.34 Hz, 1H), 7.40 (d, J=8.80 Hz, 1H), 8.19 (d, J=8.80 Hz, 1H); MS:(M+H)⁺ 226.

Step 2:

Modifications: 2.2 g2,2-difluoro-7H-1,3-dioxa-7-aza-cyclopenta[a]naphthalen-6-one used, 2.1g product obtained (87% yield).

Product:

Data: ¹H NMR (500 Hz, CDCl₃) ppm 7.51 (d, J=9.29 Hz, 1H), 7.65 (d,J=5.87 Hz, 1H), 8.22 (d, J=9.05 Hz, 1H), 8.32 (d, J=5.87 Hz, 1H); MS:(M+H)⁺ 244.

Step 3:

Modifications: 48 mg6-chloro-2,2-difluoro-1,3-dioxa-7-aza-cyclopenta[a]naphthalene and 113mg{1-[2-(1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl}-carbamicacid tert-butyl ester used, 40 mg product obtained (27% yield).

Product:

Data: ¹H NMR (400 Hz, CD₃OD) δ ppm 1.02 (s, 12H), 1.24 (m, 10H), 1.43(m, 1H), 1.88 (dd, J=8.07, 5.38 Hz, 1H), 2.32 (d, J=3.67 Hz, 2H), 2.64(d, J=13.45 Hz, 1H), 2.95 (m, 1H), 4.05 (d, J=11.49 Hz, 1H), 4.19 (d,J=9.29 Hz, 1H), 4.53 (m, 2H), 5.12 (d, J=9.78 Hz, 1H), 5.32 (s, 1H),5.77 (m, 2H), 7.34 (d, J=5.87 Hz, 1H), 7.46 (d, J=9.05 Hz, 1H), 8.11 (m,2H); MS: (M+H)⁺ 764.

Example 288 Preparation of Compound 288

Compound 288 was prepared by following Scheme 2 of Example 269 exceptthat 3-(2,2-difluoro-benzo[1,3]dioxol-5-yl)-acrylic acid was used inplace of 2-trifluormethoxycinnamic acid in step 1.

Step 1:

Modifications: 1 g 3-(2,2-difluoro-benzo[1,3]dioxol-5-yl)-acrylic acidused, 0.55 g product obtained.

Product:

Data: ¹H NMR (400 MHz, CD₃DOD) δ ppm 6.69 (d, J=7.09 Hz, 1H), 7.19 (d,J=7.09 Hz, 1H), 7.47 (s, 1H) 7.98 (s, 1H); MS: (M+H)⁺ 226.

Step 2:

Modifications: 0.5 g2,2-difluoro-6H-[1,3]dioxolo[4,5-g]isoquinolin-5-one used, 0.4 g productobtained.

Product:

Data: ¹H NMR (400 Hz, CDCl₃) δ 7.41 (s, 1H), 7.57 (d, J=5.49 Hz, 1H),7.94 (s, 1H), 8.27 (d, J=5.80 Hz, 1H); MS (M+H)⁺ 244.

Step 3:

Modifications: 48 mg5-chloro-2,2-difluoro-[1,3]dioxolo[4,5-g]isoquinoline and 112 mg{1-[2-(1-Cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl}-carbamicacid tert-butyl ester used, 30 mg product obtained.

Product:

Data: ¹H NMR (400 Hz, CD₃OD) δ ppm 1.06 (m, 12H), 1.25 (m, 10H), 1.42(m, 1H), 1.87 (dd, J=8.07, 5.62 Hz, 1H), 2.26 (m, 2H), 2.61 (dd,J=13.57, 6.97 Hz, 1H), 2.93 (m, 1H), 4.07 (dd, J=11.86, 2.81 Hz, 1H),4.22 (m, 1H), 4.40 (d, J=11.98 Hz, 1H), 4.52 (m, 1H), 5.11 (d, J=10.52Hz, 1H), 5.29 (d, J=17.12 Hz, 1H), 5.37 (s, 1H), 5.74 (m, 1H), 7.39 (s,1H), 7.56 (s, 1H), 7.63 (d, J=5.62 Hz, 1H), 7.99 (d, J=5.62 Hz, 1H).

Example 289 Preparation of Compound 289

A suspension of compound 287 (15 mg) and Pt(S)/C (5%, 5 mg) in ethylacetate (5 mL) was hydrogenated at 10 psi for 30 min. After filtration,concentration quantitatively gave 15 mg of compound 289 as a solid. ¹HNMR (400 MHz, CD₃OD) δ ppm 1.09 (m, 26H), 1.57 (m, 4H), 2.30 (m, 1H),2.61 (m, J=13.82, 7.21 Hz, 1H), 2.96 (m, 1H), 4.05 (m, J=13.94 Hz, 1H),4.19 (d, J=9.54 Hz, 1H), 4.53 (m, 2H), 5.89 (s, 1H), 7.34 (d, J=5.87 Hz,1H), 7.46 (d, J=9.05 Hz, 1H), 8.09 (d, J=5.87 Hz, 1H), 8.12 (d, J=8.80Hz, 1H); MS: (M+H)⁺ 766.

Example 290 Preparation of Compound 290

Compound 290 (15 mg, 100%) was prepared by following Scheme 3 of Example289 by using 15 mg of compound 286. Data: ¹H NMR (400 Hz, CD₃OD) δ ppm1.02 (m, 14H), 1.23 (m, 12H), 1.58 (m, 4H), 2.25 (m, 1H), 2.58 (dd,J=13.82, 7.21 Hz, 1H), 2.96 (m, 1H), 4.05 (m, J=11.25, 2.93 Hz, 1H),4.25 (d, J=9.54 Hz, 1H), 4.39 (m, 5H), 4.52 (m, J=10.03, 7.34 Hz, 1H),5.81 (s, 1H), 7.03 (d, J=9.05 Hz, 1H), 7.43 (d, J=6.11 Hz, 1H), 7.69 (d,J=9.05 Hz, 1H), 7.88 (d, J=6.11 Hz, 1H); MS: (M+H)⁺ 744.

Example 291 Preparation of Compound 291

Compound 291 (28 mg, 100%) was prepared by following Scheme 3 of Example289 by using 28 mg of compound 251. Data: ¹H NMR (400 MHz, CD₃OD) δ ppm1.01 (m, 15H), 1.26 (m, 11H), 1.37 (m, 1H), 1.58 (m, 3H), 2.25 (m, 1H),2.58 (dd, J=13.6, 7.0 Hz, 1H), 2.96 (m, 1H), 3.99 (s, 3H), 4.06 (m, 1H),4.25 (m, 1H), 4.44 (m, 1H), 4.53 (dd, J=10.3, 7.6 Hz, 1H), 5.78 (s, 1H),6.64 (d, J=9.8 Hz, 1H), 7.55 (m, 2H), 7.71 (t, J=7.3 Hz, 1H), 8.09 (d,J=8.6 Hz, 1H), 8.14 (d, J=8.1 Hz, 1H); MS: (M+Na)⁺ 738.

Example 292 Preparation of Compound 292

Compound 292

Compound 292 (16 mg, 84%) was prepared by following Scheme 3 of Example289 by using 19 mg of compound 253. ¹H NMR (400 MHz, CD₃OD) δ ppm 0.90(m, 15H), 1.15 (m, 12H), 1.48 (m, 3H), 2.18 (m, 1H), 2.51 (dd, J=13.7,6.9 Hz, 1H), 2.88 (m, 1H), 3.90 (s, 3H), 3.98 (dd, J=11.6, 3.1 Hz, 1H),4.18 (d, J=9.5 Hz, 1H), 4.36 (d, J=11.0 Hz, 1H), 4.45 (dd, J=10.2, 7.2Hz, 1H), 5.76 (s, 1H), 6.56 (d, J=9.3 Hz, 1H), 7.05 (d, J=7.6 Hz, 1H),7.34 (t, J=8.1 Hz, 1H), 7.51 (d, J=5.9 Hz, 1H), 7.65 (d, J=8.3 Hz, 1H),7.86 (d, J=6.1 Hz, 1H); MS: (M+Na)⁺ 738.

Example 293 Preparation of Compound 293

Compound 293 (7 mg, 35%) was prepared by following Scheme 3 of Example289 by using 20 mg of compound 252. ¹H NMR (400 MHz, CD₃OD) δ ppm 1.04(m, 15H), 1.27 (m, 12H), 1.58 (m, 3H), 2.27 (m, 1H), 2.60 (m, 4H), 2.96(m, 1H), 4.07 (dd, J=11.7, 2.9 Hz, 1H), 4.25 (s, 1H), 4.46 (d, J=12.0Hz, 1H), 4.54 (dd, J=10.0, 7.6 Hz, 1H), 5.85 (s, 1H), 7.39 (t, J=7.7 Hz,1H), 7.44 (d, J=5.9 Hz, 1H), 7.53 (d, J=6.9 Hz, 1H), 8.00 (d, J=6.1 Hz,1H), 8.06 (d, J=8.6 Hz, 1H); MS: (M+H)⁺ 700.

Example 294 Preparation of Compound 294

Compound 294 (14 mg, 78%) was prepared by following Scheme 3 of Example289 by using 18 mg of compound 254. ¹H NMR (400 MHz, CD₃OD) δ ppm 0.94(m, 15H), 1.13 (m, 10H), 1.20 (m, 2H), 1.50 (m, 3H), 2.21 (m, 1H), 2.53(dd, J=13.8, 7.0 Hz, 1H), 2.88 (m, 1H), 3.99 (dd, J=11.4, 2.6 Hz, 1H),4.14 (d, J=9.3 Hz, 1H), 4.45 (m, 2H), 5.79 (s, 1H), 6.53 (d, J=9.1 Hz,1H), 7.40 (t, J=8.0 Hz, 1H), 7.54 (d, J=5.9 Hz, 1H), 7.73 (d, J=7.3 Hz,1H), 8.02 (d, J=6.1 Hz, 1H), 8.10 (d, J=8.6 Hz, 1H); MS: (M+Na)⁷⁴².

Example 295 Preparation of Compound 295

Compound 295 (30 mg, 100%) was prepared by following Scheme 3 of Example289 by using 30 mg of compound 270. MS: (M+Na)⁺ 776.

Example 296 Preparation of Compound 296

Compound 296 (6.3 mg, 33%) was prepared by following Scheme 3 of Example289 by using 20 mg of compound 259. ¹H NMR (400 MHz, CD₃OD) δ ppm 1.04(m, 15H), 1.24 (m, 12H), 1.59 (m, 3H), 2.29 (m, 1H), 2.50 (s, 3H), 2.60(dd, J=13.69, 6.85 Hz, 1H), 2.97 (m, 1H), 4.09 (dd, J=11.74, 2.93 Hz,1H) 4.22 (s, 1H) 4.43 (d, J=11.74 Hz, 1H), 4.59 (dd, J=10.27, 6.85 Hz,1H), 5.87 (s, 1H), 7.30 (d, J=5.87 Hz, 1H), 7.57 (dd, J=8.31, 1.47 Hz,1H), 7.72 (d, J=8.31 Hz, 1H), 7.89 (d, J=5.87 Hz, 1H), 7.94 (s, 1H); MS:(M+H)⁺ 700.

Example 297 Preparation of Compound 297

Compound 297 (40 mg, 100%) was prepared by following Scheme 3 of Example289 by using 40 mg of compound 263. ¹H NMR (400 MHz, CD₃OD) δ ppm 0.98(m, 13H) 1.07 (m, 2H) 1.27 (m, 12H) 1.57 (m, 3H) 2.27 (m, 1H) 2.58 (dd,J=14.7, 7.1 Hz, 1H) 2.77 (s, 3H) 2.96 (m, 1H) 4.04 (m, 1H) 4.27 (m, 1H)4.42 (d, J=11.5 Hz, 1H) 4.55 (dd, J=10.6, 7.0 Hz, 1H) 5.94 (s, 1H) 6.65(d, J=9.5 Hz, 1H) 7.28 (m, 2H) 7.50 (t, J=7.6 Hz, 1H) 7.60 (d, J=7.6 Hz,1H) 7.89 (d, J=5.6 Hz, 1H); MS: (M+Na)⁺ 722.

Example 298 Preparation of Compound 298

Compound 298 (29 mg, 100%) was prepared by following Scheme 3 of Example289 by using 29 mg of compound 261. ¹H NMR (400 MHz, CD₃OD) δ ppm 0.99(m, 15H), 1.25 (m, 12H), 1.60 (m, 3H), 2.28 (m, 1H), 2.58 (m, 1H), 2.96(m, 1H), 4.02 (m, 4H), 4.22 (dd, J=8.8, 4.4 Hz, 1H), 4.48 (m, 2H), 5.83(s, 1H), 7.27 (d, J=5.6 Hz, 1H), 7.36 (d, J=8.8 Hz, 1H), 7.75 (d, J=11.3Hz, 1H), 7.90 (d, J=5.9 Hz, 1H); MS: (M+Na)⁺ 756.

Example 299 Preparation of Compound 299

Compound 299 (34 mg, 97%) was prepared by following Scheme 3 of Example289 by using 35 mg of compound 274. ¹H NMR (400 MHz, CD₃OD) δ ppm 1.01(m, 16H), 1.28 (m, 12H), 1.58 (m, 2H), 2.27 (s, 1H), 2.59 (dd, J=13.82,6.97 Hz, 1H), 2.96 (m, 1H), 4.02 (s, 3H), 4.07 (m, 1H), 4.21 (m, 1H),4.44 (m, J=11.98 Hz, 1H), 4.55 (d, J=10.27 Hz, 1H), 5.85 (s, 1H), 7.39(m, 2H), 7.95 (d, J=6.11 Hz, 1H), 8.00 (d, J=9.29 Hz, 1H); MS: (M+Na)⁺756.

Example 300 Preparation of Compound 300

Compound 300 (30 mg, 100%) was prepared by following Scheme 3 of Example289 by using 30 mg of compound 262. ¹H NMR (400 MHz, CD₃OD) δ ppm 1.27(m, 30H), 2.25 (s, 1H), 2.54 (s, 1H), 2.96 (m, 1H), 3.97 (s, 3H), 4.20(m, 3H), 4.51 (m, J=10.52, 6.85 Hz, 1H), 5.37 (s, 1H), 7.38 (s, 1H),7.62 (s, 1H), 7.65 (d, J=5.38 Hz, 1H) 8.06 (d, J=5.62 Hz, 1H); MS:(M+Na)⁺ 773.

Section G

The LC/MS method used in section G is the following:

4.6×50 mm Xterra 3 min gradient and 4 mL/min flow

Example 320 Preparation of Compound 320

Compound 320 was prepared by following Scheme 1 and Scheme 3 of above.

Step 1 (Scheme 1):

To a solution of N,N-Diethyl-4-methoxy-2-methyl-benzamide (332 mg, 1.5mmol) in THF (15 mL) at −78° C., t-BuLi (1.7 M solution in pentane, 1.3mL, 2.25 mmol) was added. The resulting red solution was stirred at −78°C. for 10 min, then 2-cyanopyridine (156 mg, 1.5 mmol) was added. Thereaction mixture was then warmed to rt and stirred for overnight. Thereaction was quenched with saturated NH₄Cl solution and extracted withethyl acetate twice. The combined organic layers were dried (MgSO₄) andconcentrated. The crude product was purified by Prep. HPLC to giveyellowish solid as TFA salt. (85 mg, 15% yield)

¹H NMR (400 MHz, CD₃OD) δ 3.91 (m, 3H), 7.09 (dd, J=9.05, 2.45 Hz, 1H),7.17 (d, J=2.45 Hz, 1H), 7.37 (s, 1H), 7.42 (m, 1H), 7.92 (m, 1H), 8.08(d, J=8.07 Hz, 1H), 8.18 (d, J=9.05 Hz, 1H), 8.65 (d, J=4.89 Hz, 1H).

LC-MS (retention time: 2.14 min.), MS m/z 253 (MH⁺).

Step 2 (Scheme 3, Step 1):

6-Methoxy-3-pyridin-2-yl-2H-isoquinolin-1-one TFA salt (85 mg, 0.232mmol) was heated under reflux with POCl₃ (3.0 mL) for 2 days. Then POCl₃was distilled off and the residue was quenched with ice. It was thenneutralized with 10 N NaOH solution and the brown solid was collected aspure product. (62 mg, 99% yield)

LC-MS (retention time: 2.063 min.), MS m/z 271 (MH⁺).

Step 3 (Scheme 3, Step 2):

To a solution of{1-[2-(1-Cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropyl-carbamoyl)-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl}-carbamicacid tert-butyl ester (82 mg, 0.148 mmol) and LaCl₃ (36 mg, 0.148 mmol)in DMF (1.5 mL), potassium t-butoxide (1.0 M solution in THF, 0.74 mL,0.74 mmol) was added at −78° C. The reaction mixture was stirred for 1hr, then 1-chloro-6-methoxy-3-pyridin-2-yl-isoquinoline (40 mg, 0.148mmol) was added. It was warmed to rt and stirred for overnight. Then itwas quenched with water and filtered. The filtrated was concentrated andthe residue was purified by Prep. HPLC to give an off-white solid asproduct (Compound 320). (23 mg, 20% yield)

¹H NMR (400 MHz, CD₃OD) δ 0.87-1.08 (m, 11H), 1.20-1.30 (m, 11H), 1.43(m, 1H), 1.87 (m, 1H), 2.22 (m, 1H), 2.35 (m, 1H), 2.69 (m, 1H), 2.93(m, 1H), 3.94 (s, 3H), 4.16 (m, 1H), 4.27 (m, 1H), 4.45 (m, 1H), 4.56(m, 1H), 5.10 (d, J=11.3 Hz, 1H), 5.27 (d, J=15.9 Hz, 1H), 5.74 (m, 1H),6.07 (s, 1H), 7.12 (d, J=7.33 Hz, 1H), 7.31 (d, J=1.96 Hz, 1H), 7.40 (m,1H), 7.94 (dd, J=7.8 Hz, 1.5 Hz, 1H), 8.11 (d, J=9.29 Hz, 1H), 8.22 (s,1H), 8.45 (d, J=8.07 Hz, 1H), 8.62 (m, 1H).

LC-MS (retention time: 2.393 min.), MS m/z 791 (MH⁺).

Example 321 Preparation of Compound 321

Condensation of ethyl bromopyruvate with ethyl thiourea in refluxingdioxane afforded the monoalkylamino thiazole as HBr salt in quantitativeyield. Alkylation of 2-ethylamino-thiazole-4-carboxylic acid ethyl esterwith EtI in DMF provided 2-diethylamino-thiazole-4-carboxylic acid ethylester.

LC/MS m/z 229 (MH⁺)

Compound 321 was prepared by following Scheme 2 and Scheme 3 above withthat 2-diethylamino-thiazole-4-carboxylic acid ethyl ester was used inthe step 1 of Scheme 2.

LC/MS (Retention time 2.76 min): m/z 868 (MH⁺).

Example 322 Preparation of Compound 322

Compound 322 was prepared by following Example 321, except that2-dimethylamino-thiazole-4-carboxylic acid ethyl ester (Preparedaccording to Scheme 5, except that methyl thiourea and methyl iodidewere used in the place of ethyl thiourea and ethyl iodide) was used inthe place of 2-dimethylamino-thiazole-4-carboxylic acid ethyl ester instep 1 of Scheme 2.

LC/MS (Retention time 2.56 min): m/z 840 (MH⁺)

Example 323 Preparation of Compound 323

Compound 323 was prepared by following Step 3 of Example 324, exceptthat 3-chloro-6-methoxy-benzo[d]isoxazole was used in the place of1-chloro-6-methoxy-3-pyridin-2-yl-isoquinoline.

MS m/z 702 (M−H)⁻

Example 324 Preparation of Compound 324

Compound 324 was prepared by following Step 3 of Example 324, exceptthat 3-chloro-benzo[d]isothiazole was used in the place of1-chloro-6-methoxy-3-pyridin-2-yl-isoquinoline.

LC/MS (Retention time 1.83 min): m/z 688 (M−H)

Example 325 Preparation of Compound 325

Compound 325 was prepared by following Scheme 1 and Scheme 3 of above.

Step 1 (Scheme 1):

To a solution of N,N-Diethyl-4-methoxy-2-methyl-benzamide (332 mg, 1.5mmol) in THF (15 mL) at −78° C., t-BuLi (1.7 M solution in pentane, 1.3mL, 2.25 mmol) was added. The resulting red solution was stirred at −78°C. for 10 min, then 4-cyanopyridine (164 mg, 1.575 mmol) was added. Thereaction mixture was then warmed to rt and stirred for overnight. Thereaction was quenched with saturated NH₄Cl solution and the yellowprecipitate was collected as pure product. (145 mg, 38% yield)

¹H NMR (CD₃OD, 400 MHz) δ 3.91 (s, 3H), 7.18 (dd, J=8.8 Hz, 2.8 Hz, 1H),7.26 (m, 2H), 8.06 (d, J=6.0 Hz, 2H), 8.16 (d, J=8.8 Hz, 1H), 8.84 (d,J=6.0 Hz, 2H).

LC-MS (retention time: 1.300 min.), MS m/z 253 (MH⁺).

Step 2 (Scheme 3, Step 1):

6-Methoxy-3-pyridin-4-yl-2H-isoquinolin-1-one (134 mg, 0.531 mmol) washeated under reflux with POCl₃ (6.0 mL) for 5 days. Then POCl₃ wasdistilled off and the residue was quenched with ice. It was thenneutralized with saturated NaHCO₃ solution and the brown solid wascollected as pure product. (125 mg, 87% yield)

¹H NMR (DMSO-d⁶, 400 MHz) δ 3.99 (s, 3H), 7.53 (dd, J=9.04 Hz, 2.44 Hz,1H), 7.59 (d, J=2.69 Hz, 1H), 8.26 (d, J=9.05 Hz, 1H), 8.30 (d, J=5.38Hz, 2H), 8.73 (s, 1H), 8.85 (d, J=6.36 Hz, 2H).

LC-MS (retention time: 2.027 min.), MS m/z 271 (MH⁺).

Step 3 (Scheme 3, Step 2):

To a solution of{1-[2-(1-Cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropyl-carbamoyl)-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl}-carbamicacid tert-butyl ester (83.5 mg, 0.15 mmol) and LaCl₃ (36.8 mg, 0.15mmol) in DMF (1.5 mL), potassium t-butoxide (1.0 M solution in THF, 0.75mL, 0.75 mmol) was added at −78° C. The reaction mixture was stirred for1 hr, then 1-chloro-6-methoxy-3-pyridin-4-yl-isoquinoline (40.6 mg, 0.15mmol) was added. It was warmed to rt and stirred for overnight. Then itwas quenched with water and filtered. The filtrated was concentrated andthe residue was purified by Prep. HPLC to give an off-white solid asproduct (Compound 325). (1.6 mg, 1.3% yield)

¹H NMR (400 MHz, CD₃OD) δ 0.90 (m, 2H), 1.02 (s, 9H), 1.17-1.31 (m,11H), 1.42 (m, 1H), 1.87 (m, 1H), 2.23 (m, 1H), 2.35 (m, 1H), 2.68 (m,1H), 2.93 (m, 1H), 3.95 (s, 3H), 4.15 (m, 1H), 4.25 (m, 1H), 4.45 (m,1H), 4.56 (m, 1H), 5.10 (d, J=10.76 Hz, 1H), 5.27 (d, J=17.61 Hz, 1H),5.74 (m, 1H), 6.06 (s, 1H), 7.14 (d, J=8.07 Hz, 1H), 7.34 (s, 1H), 8.01(s, 1H), 8.12 (d, J=8.81 Hz, 1H), 8.19 (d, J=6.12 Hz, 2H), 8.61 (d,J=5.63 Hz, 2H).

LC-MS (retention time: 2.523 min.), MS m/z 791 (MH⁺).

Example 326 Preparation of Compound 326

Compound 326 was prepared by following Scheme 1 and Scheme 4 of above.

Step 1 (Scheme 1):

To a solution of N,N-Diethyl-4-methoxy-2-methyl-benzamide (332 mg, 1.5mmol) in THF (15 mL) at −78° C., t-BuLi (1.7 M solution in pentane, 1.3mL, 2.25 mmol) was added. The resulting red solution was stirred at −78°C. for 10 min, then 4-dimethylamino benzonitrile (219 mg, 1.5 mmol) wasadded. The reaction mixture was then warmed to rt and stirred forovernight. The reaction was quenched with saturated NH₄Cl solution andthe yellow precipitate was collected and triturated with ether to givean off-white solid as pure product. (247 mg, 56% yield)

¹H NMR (DMSO-d⁶, 400 MHz) δ 2.97 (s, 6H), 3.87 (s, 3H), 6.72 (s, 1H),6.78 (d, J=8.80 Hz, 2H), 6.97 (dd, J=8.80, 2.45 Hz, 1H), 7.10 (d, J=2.45Hz, 1H), 7.65 (d, J=8.80 Hz, 2H), 8.05 (d, J=8.80 Hz, 1H), 11.11 (s,1H).

LC-MS (retention time: 2.023 min.), MS m/z 295 (MH⁺).

Step 2 (Scheme 4, Step 1):

3-(4-Dimethylamino-phenyl)-6-methoxy-2H-isoquinolin-1-one (245 mg, 0.83mmol) was heated under reflux with POCl₃ (10.0 mL) for 2 days. ThenPOCl₃ was distilled off and the residue was quenched with ice. It wasthen neutralized with 10 N NaOH solution and extracted with ethylacetate twice. The organic layers were combined and dried (MgSO₄).Evaporation of solvent gave an orange solid as product (215 mg, 83%yield)

¹H NMR (400 MHz, CD₃OD) δ 3.01 (s, 6H), 3.96 (s, 3H), 6.88 (d, J=9.05Hz, 2H), 7.20 (dd, J=9.17, 2.57 Hz, 1H), 7.28 (d, J=2.45 Hz, 1H), 7.94(s, 1H), 7.96 (d, J=9.05 Hz, 2H), 8.13 (d, J=9.29 Hz, 1H).

LC-MS (retention time: 2.543 min.), MS m/z 313 (MH⁺).

Step 3 (Scheme 4, Step 2):

A mixture of[4-(1-Chloro-6-methoxy-isoquinolin-3-yl)-phenyl]-dimethyl-amine (110 mg,0.35 mmol) and tetrabutyl phosphonium hydrogen difluoride (0.5 g) washeated at 140° C. in Smith microwave reactor for 20 min. Then it wasadded water and extracted with ethyl acetate. The organic layer wasseparated, washed with water and dried (MgSO₄). Evaporation of solventgave a brownish solid as product. (85 mg, 82% yield)

LC-MS (retention time: 2.320 min.), MS m/z 297 (MH⁺).

Step 4 (Scheme 4, Step 3):

To a solution of{1-[2-(1-Cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropyl-carbamoyl)-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl}-carbamicacid tert-butyl ester (111 mg, 0.2 mmol) and LaCl₃ (49 mg, 0.2 mmol) inDMF (2.0 mL), potassium t-butoxide (1.0 M solution in THF, 1.0 mL, 1.0mmol) was added at −78° C. The reaction mixture was stirred for 1 hr,then [4-(1-fluoro-6-methoxy-isoquinolin-3-yl)-phenyl]-dimethylamine (59mg, 0.2 mmol) was added. It was warmed to rt and stirred for overnight.Then it was quenched with water and filtered.

The filtrated was concentrated and the residue was purified by Prep.HPLC to give yellowish solid as product (Compound 326). (17.5 mg, 11%yield)

¹H NMR (400 MHz, CD₃OD) δ 0.97-1.08 (m, 11H), 1.23 (m, 2H), 1.31 (s,9H), 1.44 (m, 1H), 1.87 (m, 1H), 2.22 (m, 1H), 2.34 (m, 1H), 2.68 (m,1H), 2.93 (m, 1H), 2.99 (m, 6H), 3.91 (s, 3H), 4.17 (m, 1H), 4.29 (m,1H), 4.39 (m, 1H), 4.52 (m, 1H), 5.10 (d, J=10.76 Hz, 1H), 5.27 (d,J=17.11 Hz, 1H), 5.74 (m, 1H), 6.03 (s, 1H), 6.83 (m, 2H), 6.95 (m, 1H),7.16 (s, 1H), 7.59 (s, 1H), 8.01 (m, 3H).

LC-MS (retention time: 2.850 min.), MS m/z 834 (MH⁺).

Example 327 Preparation of Compound 327

Compound 327 was prepared by following Scheme 1 and Scheme 4 of above.

Step 1 (Scheme 1):

To a solution of N,N-Diethyl-4-methoxy-2-methyl-benzamide (332 mg, 1.5mmol) in THF (15 mL) at −78° C., t-BuLi (1.7 M solution in pentane, 1.3mL, 2.25 mmol) was added. The resulting red solution was stirred at −78°C. for 10 min, then 4-diethylamino benzonitrile (261 mg, 1.5 mmol) wasadded. The reaction mixture was then warmed to rt and stirred forovernight. The reaction was quenched with saturated NH₄Cl solution andthe yellow precipitate was collected as pure product. (215 mg, 44%yield)

¹H NMR (400 MHz, DMSO-d⁶) δ 1.12 (m, 6H), 3.39 (m, 4H), 3.87 (s, 3H),6.69 (s, 1H), 6.72 (d, J=9.05 Hz, 2H), 6.96 (dd, J=8.80, 2.45 Hz, 1H),7.09 (d, J=2.45 Hz, 1H), 7.61 (d, J=9.05 Hz, 2H), 8.04 (d, J=8.80 Hz,1H), 11.06 (s, 1H).

LC-MS (retention time: 1.883 min.), MS m/z 323 (MH⁺).

Step 2 (Scheme 4, Step 1):

3-(4-Diethylamino-phenyl)-6-methoxy-2H-isoquinolin-1-one (207 mg, 0.642mmol) was heated under reflux with POCl₃ (8.0 mL) for one day. ThenPOCl₃ was distilled off and the residue was quenched with ice. It wasthen neutralized with saturated NaHCO₃ solution and extracted with ethylacetate twice. The organic layers were combined and dried (MgSO₄).Evaporation of solvent gave a brownish solid as product. (180 mg, 82%yield)

LC-MS (retention time: 2.397 min.), MS m/z 341 (MH⁺).

Step 3 (Scheme 4, Step 2):

A mixture of[4-(1-Chloro-6-methoxy-isoquinolin-3-yl)-phenyl]-diethylamine (90 mg,0.264 mmol) and tetrabutyl phosphonium hydrogen difluoride (0.5 g) washeated at 140° C. in Smith microwave reactor for 20 min. Then it wasadded water and extracted with ethyl acetate. The organic layer wasseparated, washed with water and dried (MgSO₄). Evaporation of solventgave a yellowish oil as product. (70 mg, 82% yield)

LC-MS (retention time: 2.253 min.), MS m/z 325 (MH⁺).

Step 4 (Scheme 4, Step 3):

To a solution of{1-[2-(1-Cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropyl-carbamoyl)-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl}-carbamicacid tert-butyl ester (100 mg, 0.18 mmol) and LaCl₃ (66 mg, 0.27 mmol)in DMF (2.0 mL), potassium t-butoxide (1.0 M solution in THF, 0.9 mL,0.9 mmol) was added at −78° C. The reaction mixture was stirred for 1hr, then [4-(1-Fluoro-6-methoxy-isoquinolin-3-yl)-phenyl]-diethylamine(70 mg, 0.216 mmol) was added. It was warmed to rt and stirred forovernight. Then it was quenched with water and filtered.

The filtrated was concentrated and the residue was purified by Prep.HPLC to give white solid as product (Compound 327). (18 mg, 12% yield)

¹H NMR (400 MHz, CD₃OD) δ 0.95-1.07 (m, 11H), 1.18 (m, 6H), 1.25-1.38(m, 11H), 1.58 (m, 1H), 1.85 (m, 1H), 2.19 (m, 1H), 2.34 (m, 1H), 2.68(m, 1H), 2.92 (m, 1H), 3.42 (m, 4H), 3.90 (s, 3H), 4.16 (m, 1H), 4.28(m, 1H), 4.37 (m, 1H), 4.53 (m, 1H), 5.07 (d, J=11.0 Hz, 1H), 5.25 (d,J=17.36 Hz, 1H), 5.74 (m, 1H), 5.99 (s, 1H), 6.77 (d, J=8.8 Hz, 2H),6.94 (d, J=9.05 Hz, 1H), 7.14 (s, 1H), 7.56 (s, 1H), 7.95-8.02 (m, 3H).

LC-MS (retention time: 2.690 min.), MS m/z 862 (MH⁺).

Example 328 Preparation of Compound 328

Compound 328 was prepared by following Scheme 2 and Scheme 3 of above.

Step 1 (Scheme 2, Step 1):

To a solution of N,N-Diethyl-4-methoxy-2-methyl-benzamide (332 mg, 1.5mmol) in THF (15 mL) at −78° C., t-BuLi (1.7 M solution in pentane, 2.12mL, 3.6 mmol) was added. The resulting red solution was stirred at −78°C. for 10 min, then methyl nicotinate (206 mg, 1.5 mmol) was added. Thereaction mixture was stirred at −78° C. for 2 h. Then the reaction wasquenched with saturated NH₄Cl solution and extracted with ethyl acetatetwice. The combined organic layers were dried (MgSO₄) and concentrated.The crude product was purified by Prep. HPLC to give yellowish thick oilas TFA salt. (124 mg, 19% yield)

LC-MS (retention time: 1.740 min.), MS m/z 349 (M+Na⁺).

Step 2 (Scheme 2, Step 2):

N,N-Diethyl-4-methoxy-2-(2-oxo-2-pyridin-3-yl-ethyl)-benzamide (120 mg,0.272 mmol) was heated with ammonium acetate (1 g) for 3 hr. Then it wascooled down and added water. Extracted with ethyl acetate and theorganic layer was separated. It was then dried (MgSO₄) and concentratedto give a brownish solid as product. (65 mg, 95% yield)

¹H NMR (400 MHz, DMSO-d⁶) δ 3.89 (s, 3H), 6.93 (s, 1H), 7.10 (dd,J=8.80, 2.45 Hz, 1H), 7.19 (d, J=2.45 Hz, 1H), 7.52 (dd, J=7.46, 4.77Hz, 1H), 8.15 (m, 2H), 8.64 (dd, J=4.89, 1.47 Hz, 1H), 8.96 (d, J=1.71Hz, 1H), 11.51 (s, 1H).

LC-MS (retention time: 1.377 min.), MS m/z 253 (MH⁺).

Step 3 (Scheme 3, Step 1):

6-Methoxy-3-pyridin-3-yl-2H-isoquinolin-1-one (65 mg, 0.258 mmol) washeated under reflux with POCl₃ (2.5 mL) for 7 days. Then POCl₃ wasdistilled off and the residue was quenched with ice. It was thenneutralized with 10 N NaOH solution and extracted with ethyl acetatetwice. The combined organic layers were dried (MgSO₄) and concentratedto give yellow solid as product. (27 mg, 39% yield)

LC-MS (retention time: 2.090 min.), MS m/z 271 (MH⁺).

Step 4 (Scheme 3, Step 2):

To a solution of{1-[2-(1-Cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl}-carbamicacid tert-butyl ester (56 mg, 0.10 mmol) and LaCl₃ (25 mg, 0.10 mmol) inDMF (1.5 mL), potassium t-butoxide (1.0 M solution in THF, 0.5 mL, 0.5mmol) was added at −78° C. The reaction mixture was stirred for 1 hr,then 1-chloro-6-methoxy-3-pyridin-3-yl-isoquinoline (27 mg, 0.10 mmol)was added. It was warmed to rt and stirred for overnight. Then it wasquenched with water and filtered. The filtrated was concentrated and theresidue was purified by Prep. HPLC to give white solid as product(Compound 328). (17 mg, 21% yield)

¹H NMR (400 MHz, CD₃OD) δ 0.95 (m, 2H), 1.02 (s, 9H), 1.20-1.30 (m,11H), 1.41 (m, 1H), 1.86 (m, 1H), 2.21 (m, 1H), 2.35 (m, 1H), 2.67 (m,1H), 2.93 (m, 1H), 3.93 (s, 3H), 4.14 (m, 1H), 4.26 (m, 1H), 4.47 (d,J=11.99 Hz, 1H), 4.55 (m, 1H), 5.09 (d, J=10.02 Hz, 1H), 5.26 (d,J=17.85 Hz, 1H), 5.74 (m, 1H), 6.07 (s, 1H), 7.09 (m, 1H), 7.29 (d,J=1.96 Hz, 1H), 7.53 (m, 1H), 7.86 (s, 1H), 8.09 (d, J=9.05 Hz, 1H),8.50-8.58 (m, 2H), 9.28 (s, 1H).

LC-MS (retention time: 2.453 min.), MS m/z 791 (MH⁺).

Example 329 Preparation of Compound 329

Compound 329 was prepared by following Scheme 2 and Scheme 4 of above.

Step 1 (Scheme 2, Step 1):

To a solution of N,N-Diethyl-4-methoxy-2-methyl-benzamide (332 mg, 1.5mmol) in THF (15 mL) at −78° C., t-BuLi (1.7 M solution in pentane, 2.2mL, 3.75 mmol) was added. The resulting red solution was stirred at −78°C. for 10 min, then N,N-dimethylanthranilic acid methyl ester (269 mg,1.5 mmol) was added. The reaction mixture was stirred at −78° C. for 2h. Then the reaction was quenched with saturated NH₄Cl solution andextracted with ethyl acetate twice. The combined organic layers weredried (MgSO₄) and concentrated. The crude product was purified by Prep.HPLC to give yellowish thick oil as product. (256 mg, 46% yield)

¹H NMR (400 MHz, CD₃OD) δ 0.99-1.13 (m, 6H), 3.23-3.31 (m, 8H), 3.39 (m,2H), 3.82 (s, 3H), 4.35 (s, 2H), 6.91 (dd, J=8.44, 2.57 Hz, 1H), 6.99(d, J=2.45 Hz, 1H), 7.22 (d, J=8.56 Hz, 1H), 7.69 (t, J=7.70 Hz, 1H),7.84 (m, 1H), 7.96 (d, J=8.31 Hz, 1H), 8.18 (d, J=7.83 Hz, 1H).

LC-MS (retention time: 1.557 min.), MS m/z 369 (MH⁺).

Step 2 (Scheme 2, Step 2):

2-[2-(2-Dimethylamino-phenyl)-2-oxo-ethyl]-N,N-diethyl-4-methoxy-benzamide(250 mg, 0.678 mmol) was heated with ammonium acetate (1.5 g) for 2 hr.Then it was cooled down and added water. Extracted with ethyl acetateand the organic layer was separated. It was then dried (MgSO₄) andconcentrated to give a yellowish solid as product. (125 mg, 63% yield)

¹H NMR (400 MHz, CD₃OD) δ 2.95 (s, 6H), 3.92 (s, 3H), 6.92 (s, 1H), 7.12(dd, J=8.80, 2.45 Hz, 1H), 7.16 (d, J=2.45 Hz, 1H), 7.35 (m, 1H), 7.55(m, 2H), 7.63 (d, J=7.83 Hz, 1H), 8.20 (d, J=9.05 Hz, 1H).

LC-MS (retention time: 2.097 min.), MS m/z 295 (MH⁺).

Step 3 (Scheme 4, Step 1):

3-(2-Dimethylamino-phenyl)-6-methoxy-2H-isoquinolin-1-one (125 mg, 0.425mmol) was heated under reflux with POCl₃ (4.0 mL) for one day. ThenPOCl₃ was distilled off and the residue was quenched with ice. It wasthen neutralized with 10 N NaOH solution and extracted with ethylacetate twice. The organic layers were combined and dried (MgSO₄).Evaporation of solvent gave a brownish solid as product (82 mg, 62%yield)

LC-MS (retention time: 2.040 min.), MS m/z 313 (MH⁺).

Step 4 (Scheme 4, Step 2):

A mixture of[2-(1-Chloro-6-methoxy-isoquinolin-3-yl)-phenyl]-dimethyl-amine (82 mg,0.262 mmol) and tetrabutyl phosphonium hydrogen difluoride (1.0 g) washeated at 140° C. in Smith microwave reactor for 20 min. Then it wasadded water and extracted with ethyl acetate. The organic layer wasseparated, washed with water and dried (MgSO₄). Evaporation of solventgave the crude product which was purified by Prep. HPLC to afford ayellowish oil as product. (85 mg)

¹H NMR (400 MHz, CD₃OD) δ 3.41 (s, 6H), 4.00 (s, 3H), 7.42 (dd, J=9.05,2.45 Hz, 1H), 7.53 (s, 1H), 7.71 (m, 2H), 7.99 (m, 1H), 8.16 (m, 2H),8.31 (s, 1H).

LC-MS (retention time: 1.873 min.), MS m/z 297 (MH⁺).

Step 5 (Scheme 4, Step 3):

To a solution of{1-[2-(1-Cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropyl-carbamoyl)-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl}-carbamicacid tert-butyl ester (56 mg, 0.1 mmol) and LaCl₃ (25 mg, 0.1 mmol) inDMF (1.0 mL), potassium t-butoxide (1.0 M solution in THF, 0.5 mL, 0.5mmol) was added at −78° C. The reaction mixture was stirred for 1 hr,then [2-(1-Fluoro-6-methoxy-isoquinolin-3-yl)-phenyl]-dimethyl-amine (30mg, 0.1 mmol) was added. It was warmed to rt and stirred for overnight.Then it was quenched with water and filtered.

The filtrated was concentrated and the residue was purified by Prep.HPLC to give white solid as product (Compound 329). (4.0 mg, 5% yield)

¹H NMR (400 MHz, CD₃OD) δ 0.98-1.08 (m, 11H), 1.16-1.32 (m, 11H), 1.40(m, 1H), 1.85 (m, 1H), 2.16-2.32 (m, 2H), 2.60-2.71 (m, 7H), 2.92 (m,1H), 3.91 (s, 3H), 4.08 (m, 1H), 4.26 (m, 1H), 4.45 (m, 1H), 4.55 (m,1H), 5.10 (d, J=10.27 Hz, 1H), 5.28 (d, J=18.09 Hz, 1H), 5.74 (m, 1H),5.89 (s, 1H), 7.05 (d, J=6.85 Hz, 1H), 7.10-7.20 (m, 2H), 7.29 (m, 1H),7.63 (d, J=7.58 Hz, 1H), 7.78 (s, 1H), 8.07 (d, J=8.56 Hz, 1H).

LC-MS (retention time: 2.550 min.), MS m/z 834 (MH⁺).

Example 330 Preparation of Compound 330

Compound 330 was prepared by following Scheme 2 and Scheme 4 of above.

Step 1 (Scheme 2, Step 1):

To a solution of N,N-Diethyl-4-methoxy-2-methyl-benzamide (332 mg, 1.5mmol) in THF (15 mL) at −78° C., t-BuLi (1.7 M solution in pentane, 2.2mL, 3.75 mmol) was added. The resulting red solution was stirred at −78°C. for 10 min, then (3-dimethylamino)benzoic acid methyl ester (269 mg,1.5 mmol) was added. The reaction mixture was stirred at −78° C. for 2h. Then the reaction was quenched with saturated NH₄Cl solution andextracted with ethyl acetate twice. The combined organic layers weredried (MgSO₄) and concentrated. The crude product was purified by Prep.HPLC to give yellowish thick oil as TFA salt. (245 mg, 33% yield)

¹H NMR (400 MHz, CD₃OD) δ 1.01 (t, J=6.85 Hz, 3H), 1.09 (m, 3H), 3.11(s, 6H), 3.21 (m, 2H), 3.40 (m, 2H), 3.79 (s, 3H), 4.39 (s, 2H),6.84-6.91 (m, 2H), 7.19 (d, J=8.32 Hz, 1H), 7.35 (m, 1H), 7.49 (t,J=8.07 Hz, 1H), 7.66-7.71 (m, 2H).

LC-MS (retention time: 1.930 min.), MS m/z 369 (MH⁺).

Step 2 (Scheme 2, Step 2):

2-[2-(3-Dimethylamino-phenyl)-2-oxo-ethyl]-N,N-diethyl-4-methoxy-benzamide(240 mg, 0.497 mmol) was heated with ammonium acetate (2.0 g) for 2.5hr. Then it was cooled down and added water. A brownish solid wascollected as pure product. (95 mg, 65% yield)

¹H NMR (400 MHz, CD₃OD) δ 2.98 (s, 6H), 3.88 (s, 3H), 6.74-6.87 (m, 2H),7.01-7.07 (m, 3H), 7.18 (d, J=2.44 Hz, 1H), 7.28 (t, J=7.82 Hz, 1H),8.10 (d, J=8.80 Hz, 1H).

LC-MS (retention time: 1.773 min.), MS m/z 295 (MH⁺).

Step 3 (Scheme 4, Step 1):

3-(3-Dimethylamino-phenyl)-6-methoxy-2H-isoquinolin-1-one (92 mg, 0.312mmol) was heated under reflux with POCl₃ (3.0 mL) for 2 days. Then POCl₃was distilled off and the residue was quenched with ice. It was thenneutralized with saturated NaHCO₃ solution and extracted with ethylacetate twice. The organic layers were combined and dried (MgSO₄).Evaporation of solvent gave a brownish thick oil as product. (72 mg, 74%yield)

LC-MS (retention time: 2.297 min.), MS m/z 313 (MH⁺).

Step 4 (Scheme 4, Step 2):

A mixture of[3-(1-Chloro-6-methoxy-isoquinolin-3-yl)-phenyl]-dimethylamine (72 mg,0.23 mmol) and tetrabutyl phosphonium hydrogen difluoride (0.5 g) washeated at 140° C. in Smith microwave reactor for 20 min. Then it wasadded water and extracted with ethyl acetate. The organic layer wasseparated, washed with water and dried (MgSO₄). Evaporation of solventgave a brownish oil as product. (58 mg, 85% yield)

LC-MS (retention time: 2.193 min.), MS m/z 297 (MH⁺).

Step 5 (Scheme 4, Step 3):

To a solution of{1-[2-(1-Cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropyl-carbamoyl)-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl}-carbamicacid tert-butyl ester (86 mg, 0.155 mmol) and LaCl₃ (57 mg, 0.233 mmol)in DMF (1.5 mL), potassium t-butoxide (1.0 M solution in THF, 0.5 mL,0.5 mmol) was added at −78° C. The reaction mixture was stirred for 1hr, then [3-(1-Fluoro-6-methoxy-isoquinolin-3-yl)-phenyl]-dimethylamine(55 mg, 0.185 mmol) was added. It was warmed to rt and stirred forovernight. Then it was quenched with water and filtered.

The filtrated was concentrated and the residue was purified by Prep.HPLC to give an

off-white solid as product (Compound 330). (8.0 mg, 6% yield)

¹H NMR (400 MHz, CD₃OD) δ 0.99-1.09 (m, 11H), 1.23 (m, 2H), 1.29 (s,9H), 1.42 (m, 1H), 1.86 (m, 1H), 2.21 (m, 1H), 2.33 (m, 1H), 2.70 (m,1H), 2.93 (m, 1H), 3.00 (s, 6H), 3.92 (s, 3H), 4.14 (m, 1H), 4.29 (m,1H), 4.44-4.57 (m, 2H), 5.10 (d, J=11.00 Hz, 1H), 5.27 (d, J=16.87 Hz,1H), 5.74 (m, 1H), 6.01 (s, 1H), 6.63 (d, J=8.80 Hz, 1H), 7.03 (d,J=6.85 Hz, 1H), 7.24 (s, 1H), 7.28 (t, J=8.07 Hz, 1H), 7.45 (d, J=7.82Hz, 1H), 7.59 (s, 1H), 7.72 (s, 1H), 8.05 (d, J=8.80 Hz, 1H).

LC-MS (retention time: 2.707 min.), MS m/z 834 (MH⁺).

Example 331 Preparation of Compound 331

Compound 331 was prepared by the methods described herein.

Example 334 Preparation of Compound 334

Compound 334 was prepared in the following manner:

Step 1

To a solution of Boc-cis-HYP-OMe (122.6 mg, 0.5 mmol) in THF (15 mL) at0° C., triphenylphosphine (196.7 mg, 0.75 mmol) andbenzo[d]isoxazol-3-ol (81 mg, 0.6 mmol) were added. Then DEAD (0.118 mL,0.75 mmol) was added. The reaction mixture was warmed to rt. and stirredfor 3 hr. Then solvent was evaporated and the residue was purified byPrep. HPLC to give a colorless thick oil. (117 mg, 54% yield)

¹H NMR (400 MHz, CD₃OD) δ 1.41 (m, 9H), 2.38 (m, 1H), 2.75 (m, 1H), 3.75(m, 3H), 3.81 (m, 1H), 3.90 (m, 1H), 4.47 (m, 1H), 5.44 (m, 1H), 7.31(t, J=7.46 Hz, 1H), 7.47 (d, J=8.56 Hz, 1H), 7.59 (t, J=7.83 Hz, 1H),7.66 (d, J=8.07 Hz, 1H).

LC-MS (retention time: 2.65 min.), MS m/z 363 (MH⁺).

Some of the coupling product (85 mg, 0.235 mmol) was then dissolved in4N HCl in dioxane (1.5 mL) and stirred for 3 hr. Evaporation of solventgave a yellowing oil as HCl salt. (85 mg, >100% yield)

LC-MS (retention time: 1.327 min.), MS m/z 263 (MH⁺).

Step 2:

To a solution of 4-(Benzo[d]isoxazol-3-yloxy)-pyrrolidine-2-carboxylicacid methyl ester hydrochloride salt (85 mg, 0.285 mmol) in CH₃CN (10mL) was added N-boc-L-t-leucine (99 mg, 0.427 mmol), DIEA (0.25 mL,1.425 mmol) and the coupling reagent HOBt (65 mg, 0.427 mmol) and HBTU(162 mg, 0.427 mmol). The solution was stirred at rt. overnight. Then itwas concentrated, washed with water and extracted with ethyl acetatetwice. The combined organic layers were washed with brine, dried overMgSO₄ and concentrated. It was then purified by Prep. HPLC column togive a colorless thick oil as product. (63 mg, 46% yield)

1H NMR (400 MHz, CD₃OD) δ 1.01 (s, 9H), 1.17 (s, 9H), 2.34 (m, 1H), 2.78(dd, J=14.13, 7.83 Hz, 1H), 3.72 (s, 3H), 4.00 (dd, J=12.22, 3.42 Hz,1H), 4.19 (s, 1H), 4.57 (d, J=12.23 Hz, 1H), 4.68 (m, 1H), 5.51 (m, 1H),7.27 (m, 1H), 7.47 (d, J=8.56 Hz, 1H), 7.57 (m, 1H), 7.63 (d, J=8.07 Hz,1H).

LC-MS (retention time: 2.737 min.), MS m/z 498 (M+Na⁺).

Step 3:

To a solution of4-(Benzo[d]isoxazol-3-yloxy)-1-(2-tert-butoxycarbonylamino-3,3-dimethyl-butyryl)-pyrrolidine-2-carboxylicacid methyl ester (63 mg, 0.132 mmol) in THF (3.5 mL), methanol (2.0 mL)and water (0.5 mL) mixture, lithium hydroxide monohydrate (83 mg, 1.89mmol) was added. The reaction mixture was stirred at rt. for overnight.Then it was acidified with 1N HCl solution to pH=3 to 5 andconcentrated. Extracted with ethyl acetate (2×30 mL) and the organiclayers were combined and dried (MgSO₄). Evaporation of solvent gave ayellowish oil to carry on. (61 mg, 100% yield)

To a solution of above compound (61 mg, 0.132 mmol) in CH₃CN (8 mL) wasadded (1R,2S)(1-cyclopropanesulfonyl-aminocarbonyl-2-vinyl-cyclo-propyl)-carbamicacid hydrochloride (42 mg, 0.158 mmol), DIEA (0.115 mL, 0.66 mmol) andthe coupling reagent HOBt (30 mg, 0.198 mmol) and HBTU (75 mg, 0.198mmol). The solution was stirred at rt. overnight. Then it wasconcentrated, washed with water and extracted with ethyl acetate twice.The combined organic layers were washed with brine, dried over MgSO₄ andconcentrated. It was then purified by Prep. HPLC column to give a yellowfilm as final product (Compound 334). (24 mg, 27% yield)

1H NMR (400 MHz, CD₃OD) δ 1.01 (s, 9H), 1.05 (m, 2H), 1.12-1.26 (m,11H), 1.43 (m, 1H), 1.86 (dd, J=8.07, 5.38 Hz, 1H), 2.17-2.33 (m, 2H),2.67 (dd, J=12.96, 5.87 Hz, 1H), 2.93 (m, 1H), 4.05 (m, 1H), 4.22 (m,1H), 4.49 (m, 2H), 5.11 (d, J=10.21 Hz, 1H), 5.29 (d, J=17.12 Hz, 1H),5.55 (s, 1H), 5.74 (m, 1H), 7.29 (m, 1H), 7.48 (d, J=8.32 Hz, 1H),7.54-7.64 (m, 2H).

LC-MS (retention time: 2.767 min.), MS m/z 696 (M+Na).

Example 335 Preparation of Compound 335

Step 1: (Scheme 1, step 1)

To a solution of (2S,4R) 4-hydroxy-pyrrolidine-1,2-dicarboxylic acid1-tert-butyl ester (0.25 g, 1.08 mmol) in CH₃CN (10 mL) was added(1R,2S)(1-cyclopropanesulfonyl-aminocarbonyl-2-vinyl-cyclo-propyl)-carbamicacid hydrochloride (0.346 g, 1.30 mmol), DIEA (0.94 mL, 5.41 mmol) andthe coupling reagent HOBt (0.248 g, 1.62 mmol) and HBTU (0.615 mg, 1.62mmol). The solution was stirred at rt. overnight. Then it wasconcentrated, washed with water and extracted with ethyl acetate twice.The combined organic layers were washed with brine, dried over MgSO₄ andconcentrated to give yellow oil. It was purified by Prep. HPLC column togive a colorless thick oil which was then dissolved in 4N HCl in dioxane(5 mL). The reaction mixture was stirred at rt. for overnight.Evaporation of solvent gave white solid as product to carry on. (200 mg,49% yield)

LC-MS (retention time: 0.647 min.), MS m/z 344 (MH⁺).

Step 2: (Scheme 1, Step 2)

To a solution of above compound (200 mg, 0.527 mmol) in CH₃CN (10 mL)was added 2-methoxycarbonylamino-3,3-dimethyl-butyric acid (0.15 g, 0.79mmol), DIEA (0.46 mL, 2.63 mmol) and the coupling reagent HOBt (0.121 g,0.79 mmol) and HBTU (0.30 g, 0.79 mmol). The solution was stirred at rt.overnight. Then it was concentrated, washed with water and extractedwith ethyl acetate twice. The combined organic layers were washed withbrine, dried over MgSO₄ and concentrated to give yellowish oil. It waspurified by Prep. HPLC column to give white solid as final product(intermediate 2). (145 mg, 54% yield)

¹H NMR (CD₃OD, 500 MHz) δ 0.99-1.10 (m, 11H), 1.24 (m, 2H), 1.41 (dd,J=9.5, 5.5 Hz, 1H), 1.87 (dd, J=7.9, 5.5 Hz, 1H), 1.97 (m, 1H), 2.13 (m,1H), 2.24 (m, 1H), 2.93 (m, 1H), 3.65 (s, 3H), 3.77-3.88 (m, 2H),4.33-4.39 (m, 2H), 4.49 (m, br, 1H), 5.13 (d, J=10.4 Hz, 1H), 5.31 (d,J=17.1 Hz, 1H), 5.76 (m, 1H).

LC-MS (retention time: 1.590 min.), MS m/z 515 (MH⁺).

Step 3: (Scheme 2)

To a solution of 2,4-dichloropyrimidine (149 mg, 1 mmol) in THF (5 mL),tetrakis(triphenylphosphine) palladium (23 mg, 2 mol %) and 0.5Msolution of phenylzinc bromide (2.1 mL, 1.05 mmol) in THF were added.The reaction mixture was stirred at 50° C. for overnight. Then it wasadded saturated ammonium chloride solution and extracted with EtOActwice. The organic layers were combined, washed with water and dried(MgSO₄). Evaporation of solvent gave a yellow residue which was purifiedby Prep. HPLC to afford a yellowish oil as 2-chloro-4-phenyl-pyrimidineto carry on.

To a solution of intermediate 2 (20 mg, 0.039 mmol) in DMF (3 mL), NaH(3.9 mg of 60% dispersion in mineral oil, 0.0975 mmol) was added at 0°C. The reaction mixture was then warmed to rt. and stirred for 1 hr.Then 2-chloro-4-phenyl-pyrimidine prepared above (18 mg as crude) wasadded. The reaction mixture was stirred at rt. for overnight. It wasthen quenched with water and extracted with EtOAc. The organic layer wasseparated, washed with brine and dried (MgSO₄). Evaporation of solventgave yellowish oil which was then purified by Prep. HPLC to give a thickcolorless oil as final product (Compound 335) as TFA salt. (5.5 mg, 18%yield)

¹H NMR (CD₃OD, 300 MHz) δ 0.92-1.12 (m, 11H), 1.25 (m, 2H), 1.44 (dd,J=9.2, 5.5 Hz, 1H), 1.89 (dd, J=8.1, 5.5 Hz, 1H), 2.17-2.37 (m, 2H),2.57 (m, 1H), 2.95 (m, 1H), 3.52 (s, 3H), 4.14 (m, 1H), 4.24-4.38 (m,2H), 4.51 (m, 1H), 5.13 (d, J=10.2 Hz, 1H), 5.31 (d, J=17.2 Hz, 1H),5.77 (m, 1H), 5.86 (s, 1H), 7.48-7.60 (m, 3H), 7.66 (d, J=5.3 Hz, 1H),8.18 (m, 2H), 8.60 (d, J=5.1 Hz, 1H).

LC-MS (retention time: 1.947 min.), MS m/z 669 (MH⁺).

Example 336 Preparation of Compound 336

Step 1:

To a solution of 2,4-dichloropyrimidine (149 mg, 1 mmol) in THF (5 mL),tetrakis(triphenylphosphine) palladium (58 mg, 5 mol %) and 0.5Msolution of 2-pyridinylzinc bromide (2.4 mL, 1.2 mmol) in THF wereadded. The reaction mixture was stirred at 50° C. for overnight. Then itwas added saturated ammonium chloride solution and extracted with EtOActwice. The organic layers were combined, washed with water and dried(MgSO₄). Evaporation of solvent gave a yellow residue which was purifiedby Prep. HPLC to afford a yellowish oil as product. (11 mg, 3.6% yield)

¹H NMR (500 MHz, CD₃OD) δ 7.61 (m, 1H), 8.07 (m, 1H), 8.36 (d, J=5.19Hz, 1H), 8.50 (d, J=7.94 Hz, 1H), 8.75 (d, J=3.97 Hz, 1H), 8.82 (d,J=5.19 Hz, 1H).

LC-MS (retention time: 1.440 min.), MS m/z 192 (MH⁺).

Step 2:

To a solution of intermediate 2 from Example 335 (15 mg, 0.029 mmol) inDMF (3 mL), NaH (1.75 mg of 60% dispersion in mineral oil, 0.0728 mmol)was added at 0° C. The reaction mixture was then warmed to rt. andstirred for 1 hr. Then 2-Chloro-4-pyridin-2-yl-pyrimidine (9.5 mg,0.0311 mmol) was added. The reaction mixture was stirred at rt. forovernight. It was then quenched with water and extracted with EtOAc. Theorganic layer was separated, washed with brine and dried (MgSO₄).Evaporation of solvent gave yellowish oil which was then purified byPrep. HPLC to give a yellowish film as final product (Compound 336) asTFA salt. (3.5 mg, 15% yield)

¹H NMR (CD₃OD, 500 MHz) δ 1.03 (s, 9H), 1.08 (m, 2H), 1.24 (m, 2H), 1.43(dd, J=9.77, 5.50 Hz, 1H), 1.89 (m, 1H), 2.24 (m, 1H), 2.31 (m, 1H),2.57 (m, 1H), 2.95 (m, 1H), 3.50 (s, 3H), 4.13 (m, 1H), 4.29 (s, 1H),4.36 (d, J=11.91 Hz, 1H), 4.52 (m, 1H), 5.13 (d, J=10.08 Hz, 1H), 5.31(d, J=16.79 Hz, 1H), 5.76 (m, 1H), 5.88 (m, 1H), 7.64 (m, 1H), 8.06-8.13(m, 2H), 8.54 (d, J=7.93 Hz, 1H), 8.73-8.76 (m, 2H).

LC-MS (retention time: 1.787 min.), MS m/z 670 (MH⁺).

Example 337 Preparation of Compound 337

Step 1:

To a solution of 2,4-dichloropyrimidine (149 mg, 1 mmol) in DMF (5 mL),dichloro bis(triphenylphosphine) palladium (II) (35 mg, 5 mol %) and2-(tributylstannyl)thiophene (0.38 mL, 1.2 mmol) were added. Thereaction mixture was heated at 70° C. for 3 hr. Then it was addedsaturated KF solution in methanol (20 mL) and stirred at rt for 4 hr.The reaction mixture was concentrated with a small amount of silica geland the residue was filtered through filter paper and washed with EtOAc.The filtrate was then concentrated and the residue was purified by Prep.HPLC to afford an off-white solid as product. (110 mg, 35% yield)

¹H NMR (400 MHz, CD₃OD) δ 7.20 (dd, J=5.01, 3.79 Hz, 1H), 7.74 (dd,J=5.01, 1.10 Hz, 1H), 7.77 (d, J=5.38 Hz, 1H), 7.98 (dd, J=3.79, 1.10Hz, 1H), 8.55 (d, J=5.38 Hz, 1H).

LC-MS (retention time: 1.453 min.), MS m/z 197 (MH⁺).

Step 2:

To a solution of intermediate 2 from Example 335 (20 mg, 0.039 mmol) inDMF (3 mL), NaH (7.8 mg of 60% dispersion in mineral oil, 0.195 mmol)was added at 0° C. The reaction mixture was then warmed to rt. andstirred for 1 hr. Then 2-Chloro-4-thiophen-2-yl-pyrimidine (16.9 mg,0.0544 mmol) was added. The reaction mixture was stirred at rt. forovernight. It was then quenched with water and extracted with EtOAc. Theorganic layer was separated, washed with brine and dried (MgSO₄).

Evaporation of solvent gave yellowish oil which was then purified byPrep. HPLC to give two products (Compound 337 and intermediate 3).

Compound 337: (yellowish film, 3.0 mg, 11% yield))

¹H NMR (CD₃OD, 500 MHz) δ 0.98-1.07 (m, 11H), 1.22 (m, 2H), 1.41 (dd,J=9.54, 5.62 Hz, 1H), 1.86 (dd, J=8.32, 5.63 Hz, 1H), 2.19-2.31 (m, 2H),2.52 (m, 1H), 2.92 (m, 1H), 3.50 (s, 3H), 4.09 (m, 1H), 4.25-4.32 (m,2H), 4.47 (dd, J=10.03, 7.34 Hz, 1H), 5.11 (dd, J=10.27, 1.71 Hz, 1H),5.28 (dd, J=17.11, 1.46 Hz, 1H), 5.69-5.79 (m, 2H), 7.20 (dd, J=4.89,3.66 Hz, 1H), 7.51 (d, J=5.38 Hz, 1H), 7.70 (d, J=4.89 Hz, 1H), 7.95 (d,J=3.67 Hz, 1H), 8.54 (d, J=5.14 Hz, 1H).

LC-MS (retention time: 1.787 min.), MS m/z 696 (M+Na⁺).

Intermediate 3: (10 mg, 35% yield)

LC-MS (retention time: 1.477 min.), MS m/z 617 (MH⁺).

Example 338 Preparation of Compound 338

To a solution of1-(2-Amino-3,3-dimethyl-butyryl)-4-(4-thiophen-2-yl-pyrimidin-2-yloxy)-pyrrolidine-2-carboxylicacid (1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropyl)-amide (10mg, 0.0137 mmol) in CH₃CN (5 mL) was added cyclopropylacetic acid (2.1mg, 0.0205 mmol), DIEA (0.012 mL, 0.742 mmol) and the coupling reagentHOBt (3.1 g, 0.0205 mmol) and HBTU (7.8 mg, 0.0205 mmol). The solutionwas stirred at rt. overnight. Then it was concentrated, washed withwater and extracted with ethyl acetate twice. The combined organiclayers were washed with brine, dried over MgSO₄ and concentrated to giveyellow oil. It was purified by Prep. HPLC column to give a yellowishfilm as TFA salt (Compound 338). (4.6 mg, 41% yield)

¹H NMR (CD₃OD, 400 MHz) δ 0.12 (m, 2H), 0.48 (m, 2H), 0.90 (m, 1H),1.01-1.09 (m, 11H), 1.23 (m, 2H), 1.43 (dd, J=9.29, 5.38 Hz, 1H), 1.87(dd, J=8.31, 5.62 Hz, 1H), 2.06 (m, 2H), 2.19-2.31 (m, 2H), 2.52 (dd,J=13.45, 6.85 Hz, 1H), 2.93 (m, 1H), 4.12 (dd, J=11.98, 3.91 Hz, 1H),4.27 (d, J=11.74 Hz, 1H), 4.47 (dd, J=10.27, 6.85 Hz, 1H), 4.63 (s, 1H),5.11 (dd, J=10.27, 1.47 Hz, 1H), 5.28 (dd, J=17.12, 1.47 Hz, 1H),5.71-5.80 (m, 2H), 7.20 (dd, J=4.89, 3.67 Hz, 1H), 7.51 (d, J=5.38 Hz,1H), 7.70 (d, J=5.20 Hz, 1H), 7.95 (d, J=3.67 Hz, 1H), 8.48 (d, J=5.13Hz, 1H).

LC-MS (retention time: 1.833 min.), MS m/z 699 (MH⁺).

Example 339 Preparation of Compound 339

Compound 342 was prepared by following Schemes of Example 337 andExample 338, except that 2-(tributylstannyl)furan was used in the placeof 2-(tributylstannyl)thiophene in the Step 1 of Example 337.

Step 1

To a solution of 2,4-dichloropyrimidine (149 mg, 1 mmol) in DMF (5 mL),dichloro bis(triphenylphosphine) palladium (II) (35 mg, 5 mol %) and2-(tributylstannyl)furan (0.35 mL, 1.1 mmol) were added. The reactionmixture was heated at 70° C. for 3 hr. Then it was added saturated KFsolution in methanol (20 mL) and stirred at rt for 4 hr. The reactionmixture was concentrated with a small amount of silica gel and theresidue was filtered through filter paper and washed with EtOAc. Thefiltrate was then concentrated and the residue was purified by Prep.HPLC to afford a brownish solid as product. (80 mg, 27% yield)

¹H NMR (400 MHz, CD₃OD) δ 6.68 (dd, J=3.67, 1.71 Hz, 1H), 7.42 (d,J=3.67 Hz, 1H), 7.67 (d, J=5.13 Hz, 1H), 7.30 (d, J=1.71 Hz, 1H), 8.62(d, J=5.14 Hz, 1H).

LC-MS (retention time: 1.233 min.), MS m/z 181 (MH⁺).

Step 2

To a solution of{1-[2-(1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropyl-carbamoyl)-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl}-carbamicacid methyl ester (20 mg, 0.039 mmol) in DMF (3 mL), NaH (7.8 mg of 60%dispersion in mineral oil, 0.195 mmol) was added at 0° C. The reactionmixture was then warmed to rt. and stirred for 1 hr. Then2-Chloro-4-thiophen-2-yl-pyrimidine (16.0 mg, 0.0544 mmol) was added.The reaction mixture was stirred at rt. for overnight. It was thenquenched with water and extracted with EtOAc. The organic layer wasseparated, washed with brine and dried (MgSO₄). Evaporation of solventgave yellowish oil which was then purified by Prep. HPLC to give debocedcoupling product. (3 mg, 11% yield)

LC-MS (retention time: 1.420 min.), MS m/z 601 (MH⁺).

Step 3

To a solution of1-(2-Amino-3,3-dimethyl-butyryl)-4-(4-furan-2-yl-pyrimidin-2-yloxy)-pyrrolidine-2-carboxylicacid (1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropyl)-amide (3mg, 0.0042 mmol) in CH₃CN (5 mL) was added cyclopropylacetic acid (0.6mg, 0.0063 mmol), DIEA (0.004 mL, 0.021 mmol) and the coupling reagentHOBt (1.0 g, 0.0063 mmol) and HBTU (2.4 mg, 0.0063 mmol). The solutionwas stirred at rt. overnight. Then it was concentrated, washed withwater and extracted with ethyl acetate twice. The combined organiclayers were washed with brine, dried over MgSO₄ and concentrated to giveyellow oil. It was purified by Prep. HPLC column to give a yellowishfilm as TFA salt (Compound 339). (1.0 mg, 30% yield)

¹H NMR (CD₃OD, 400 MHz) δ 0.12 (m, 2H), 0.48 (m, 2H), 0.90 (m, 1H),0.99-1.09 (m, 11H), 1.23 (m, 2H), 1.43 (dd, J=9.05, 5.31 Hz, 1H), 1.87(m, 1H), 2.05 (m, 2H), 2.19-2.29 (m, 2H), 2.50 (m, 1H), 2.93 (m, 1H),4.10 (dd, J=12.23, 3.91 Hz, 1H), 4.25 (d, J=11.99 Hz, 1H), 4.47 (dd,J=10.52, 7.09 Hz, 1H), 4.63 (s, 1H), 5.11 (dd, J=10.52, 1.71 Hz, 1H),5.29 (dd, J=17.12, 1.47 Hz, 1H), 5.71-5.79 (m, 2H), 6.65 (dd, J=3.67,1.96 Hz, 1H), 7.38 (d, J=3.67 Hz, 1H), 7.40 (d, J=5.38 Hz, 1H), 7.76 (m,1H), 8.54 (d, J=5.38 Hz, 1H).

LC-MS (retention time: 1.790 min.), MS m/z 683 (MH⁺).

Example 340 Preparation of Compound 340

To a solution of 2,4-dichloropyrimidine (149 mg, 1 mmol) in DMF (5 mL),dichloro bis(triphenylphosphine) palladium (II) (35 mg, 5 mol %) and2-(tributylstannyl)thiazole (412 mg, 1.1 mmol) were added. The reactionmixture was heated at 80° C. for 3 hr. Then it was added saturated KFsolution in methanol (20 mL) and stirred at rt for 4 hr. The reactionmixture was concentrated with a small amount of silica gel and theresidue was filtered through filter paper and washed with EtOAc.

The filtrate was then concentrated and the residue was purified by Prep.HPLC to afford a brownish solid as product. (9 mg, 3% yield)

LC-MS (retention time: 1.320 min.), MS m/z 198 (MH⁺).

Step 2:

To a solution of1-[2-(2-Cyclopropyl-acetylamino)-3,3-dimethyl-butyryl]-4-hydroxy-pyrrolidine-2-carboxylicacid (1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropyl)-amide(12.5 mg, 0.0232 mmol) in DMF (3 mL), NaH (3.7 mg of 60% dispersion inmineral oil, 0.0.0928 mmol) was added at 0° C. The reaction mixture wasthen warmed to rt. and stirred for 1 hr. Then2-Chloro-4-thiazole-2-yl-pyrimidine (9.0 mg, 0.0289 mmol) was added. Thereaction mixture was stirred at rt. for overnight. It was then quenchedwith water and extracted with EtOAc. The organic layer was separated,washed with brine and dried (MgSO₄). Evaporation of solvent gave crudeproduct which was then purified by Prep. HPLC to give white solid asfinal product (Compound 340). (2.8 mg, 17% yield)

¹H NMR (CD₃OD, 400 MHz) δ 0.12 (m, 2H), 0.47 (m, 2H), 0.89 (m, 1H),1.00-1.09 (m, 11H), 1.22 (m, 2H), 1.44 (dd, J=9.54, 5.38 Hz, 1H), 1.87(dd, J=8.07, 5.38 Hz, 1H), 2.06 (m, 2H), 2.20-2.32 (m, 2H), 2.52 (dd,J=13.70, 6.85 Hz, 1H), 2.93 (m, 1H), 4.13 (dd, J=11.98, 3.91 Hz, 1H),4.30 (d, J=11.98 Hz, 1H), 4.48 (dd, J=10.51, 7.09 Hz, 1H), 4.63 (d,J=9.54 Hz, 1H), 5.11 (d, J=10.51 Hz, 1H), 5.29 (d, J=17.12 Hz, 1H),5.73-5.80 (m, 2H), 7.81 (d, J=5.14 Hz, 1H), 7.84 (d, J=3.18 Hz, 1H),8.03 (d, J=2.93 Hz, 1H), 8.68 (d, J=5.13 Hz, 1H).

LC-MS (retention time: 1.710 min.), MS m/z 700 (MH⁺).

Example 341 Preparation of Compound 341

Step 1 (Scheme 1, Step 1):

To a solution of Boc-HYP-OH (1.0 g, 4.324 mmol) in DMF (20 mL), NaH(0.38 g of 60% dispersion in mineral oil, 9.513 mmol) was added at 0° C.The reaction mixture was stirred for 1 hr. Then 2,4-dichloropyrimidine(0.709 g, 0.0289 mmol) was added. The reaction mixture was warmed to rtand stirred for overnight. It was then quenched with 1N HCl solution andextracted with EtOAc. The organic layer was separated, washed with brineand dried (MgSO₄). Evaporation of solvent gave crude product which wasthen purified by Prep. HPLC to give colorless oil as product. (0.4 g,27% yield)

¹H NMR (CD₃OD, 300 MHz) δ 1.13 (m, 9H), 2.37 (m, 1H), 2.62 (m, 1H),3.70-3.84 (m, 2H), 4.38 (m, 1H), 5.65 (m, 1H), 6.88 (d, J=5.86 Hz, 1H),8.37 (d, J=5.86 Hz, 1H).

LC-MS (retention time: 1.370 min.), MS m/z 344 (MH⁺).

Step 2: (Scheme 1, Step 2)

To a solution of (2S,4R)4-(2-Chloro-pyrimidin-4-yloxy)-pyrrolidine-1,2-dicarboxylic acid1-tert-butyl ester (0.34 g, 0.99 mmol) in CH₃CN (20 mL) was added(1R,2S)/(1S,2R)(1-cyclopropanesulfonyl-aminocarbonyl-2-vinyl-cyclo-propyl)-carbamicacid (0.511 g, 1.48 mmol), DIEA (0.86 mL, 4.95 mmol) and the couplingreagent HOBt (0.226 g, 1.48 mmol) and HBTU (0.561 g, 1.48 mmol). Thesolution was stirred at rt. overnight. Then it was concentrated, washedwith water and extracted with ethyl acetate twice. The combined organiclayers were washed with brine, dried over MgSO₄ and concentrated. It wasthen purified by Prep. HPLC column to give a yellow solid (intermediate4). (0.33 g, 41% yield)

¹H NMR (CD₃OD, 300 MHz) δ diasteoromer mixture.

LC-MS (retention time: 2.907 min.), MS m/z 655 (MH⁺).

Step 3: (Scheme 2, Step 1)

To a solution of intermediate 4 (50 mg, 0.061 mmol) in CH₂Cl₂ (2.5 mL),1,2,3,4-tetrahydroisoquinoline (0.011 mL, 0.0915 mmol) and Et₃N (0.021mL, 0.153 mmol) were added. The reaction mixture was stirred at rt forovernight and at 40° C. for 1 day. The solvent was stripped and theresidue was purified by Prep. HPLC to give a colorless oil. It was thendissolved in 4N HCl in dioxane (1 mL) and stirred for overnight.Evaporation of solvent gave a colorless oil as hydrochloride salt. (20mg, 52% yield)

LC-MS (retention time: 1.160 min.), MS m/z 553 (MH⁺).

Step 4: (Scheme 2, Step 2)

To a solution of4-[2-(3,4-Dihydro-1H-isoquinolin-2-yl)-pyrimidin-4-yloxy]-pyrrolidine-2-carboxylicacid (1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropyl)-amidehydrochloride (20 mg, 0.032 mmol) in CH₃CN (5 mL) was added2-methoxycarbonylamino-3,3-dimethyl-butyric acid (9.1 mg, 0.048 mmol),DIEA (0.028 mL, 0.16 mmol) and the coupling reagent HOBt (7.3 mg, 0.048mmol) and HBTU (18.2 mg, 0.048 mmol). The solution was stirred at rt.overnight. Then it was concentrated, washed with water and extractedwith ethyl acetate twice. The combined organic layers were washed withbrine, dried over MgSO₄ and concentrated to give yellowish oil. It waspurified by Prep. HPLC column to give a colorless oil as TFA salt(Compound 341). (16 mg, 60% yield)

¹H NMR (CD₃OD, 500 MHz) δ 0.98-1.06 (m, 13H), 1.13 (m, 1H), 1.22-1.32(m, 1H), 1.35-1.44 (m, 1H), 1.82 (dd, J=8.24, 5.19 Hz, 0.5H), 1.90 (dd,J=8.24, 5.49 Hz, 0.5H), 2.26 (m, 1H), 2.32-2.43 (m, 1H), 2.56 (m, 1H),2.96 (m, 1H), 3.11 (m, br, 2H), 3.56 (s, 3H), 4.14 (m, 1H), 4.21 (m,1H), 4.38 (m, 1H), 4.47 (m, 1H), 5.15 (m, 1H), 5.31 (m, 1H), 5.75 (m,1H), 5.94 (s, 1H), 6.47 (d, J=7.02 Hz, 1H), 7.29 (s, 4H), 7.49 (m, 1H),7.56 (m, 1H), 7.74 (d, J=8.24 Hz, 1H), 7.88 (d, J=8.24 Hz, 1H), 8.11 (d,J=7.02 Hz, 1H).

LC-MS (retention time: 1.517 min.), MS m/z 724 (MH⁺).

Example 342 Preparation of Compound 342

Compound 342w as prepared by following Scheme 2 of Example 341, exceptthat isoindoline was used in the place of 1,2,3,4-tetrahydroisoquinolinein step 1 of scheme 2.

Step 1:

To a solution of intermediate 4 from Example 341 (50 mg, 0.061 mmol) inCH₂Cl₂ (2.5 mL), isoindoline (0.013 mL, 0.115 mmol) and Et₃N (0.026 mL,0.09 mmol) were added. The reaction mixture was stirred at rt for 2days. The solvent was stripped and the residue was purified by Prep.HPLC to give a colorless oil. It was then dissolved in 4N HCl in dioxane(1 mL) and stirred for overnight. Evaporation of solvent gave crudeproduct which was purified by Prep.HPLC again to afford yellowish solidas TFA salt. (8.5 mg, 14% yield)

LC-MS (retention time: 1.860 min.), MS m/z 539 (MH⁺).

Step 2:

To a solution of4-[2-(1,3-Dihydro-isoindol-2-yl)-pyrimidin-4-yloxy]-pyrrolidine-2-carboxylicacid (1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropyl)-amidehydrochloride (8.5 mg, 0.0104 mmol) in CH₃CN (5 mL) was added2-methoxycarbonylamino-3,3-dimethyl-butyric acid (3.0 mg, 0.0156 mmol),DIEA (0.009 mL, 0.052 mmol) and the coupling reagent HOBt (2.4 mg,0.0156 mmol) and HBTU (5.9 mg, 0.0156 mmol). The solution was stirred atrt. overnight. Then it was concentrated, washed with water and extractedwith ethyl acetate twice. The combined organic layers were washed withbrine, dried over MgSO₄ and concentrated to give yellowish oil. It waspurified by Prep. HPLC column to give a colorless oil as TFA salt(Compound 342). (3 mg, 35% yield)

¹H NMR (CD₃OD, 300 MHz) δ diasteoromer mixture.

LC-MS (retention time: 2.547 min.), MS m/z 710 (MH⁺).

Example 343 Preparation of Compound 343

Compound 342 was prepared by following Scheme 2 of Example 341, exceptthat morpholine was used in the place of 1,2,3,4-tetrahydroisoquinolinein step 1 of scheme 2.

Step 1:

To a solution of intermediate 4 from Example 341 (50 mg, 0.061 mmol) inCH₂Cl₂ (2.5 mL), morpholine (0.008 mL, 0.0915 mmol) and Et₃N (0.021 mL,0.153 mmol) were added. The reaction mixture was stirred at rt forovernight and at 40° C. for 1 day. The solvent was stripped and theresidue was purified by Prep. HPLC to give a colorless oil. It was thendissolved in 4N HCl in dioxane (1 mL) and stirred for overnight.Evaporation of solvent gave a colorless oil as hydrochloride salt. (12.6mg, 36% yield)

LC-MS (retention time: 0.810 min.), MS m/z 507 (MH⁺).

Step 2:

To a solution of4-(2-Morpholin-4-yl-pyrimidin-4-yloxy)-pyrrolidine-2-carboxylic acid(1-cyclopropanesulfonylaminocarbonyl-2-vinyl-cyclopropyl)-amidehydrochloride (12.6 mg, 0.0217 mmol) in CH₃CN (5 mL) was added2-methoxycarbonylamino-3,3-dimethyl-butyric acid (6.2 mg, 0.0326 mmol),DIEA (0.019 mL, 0.1085 mmol) and the coupling reagent HOBt (5.0 mg,0.0326 mmol) and HBTU (12.4 mg, 0.0326 mmol). The solution was stirredat rt. overnight. Then it was concentrated, washed with water andextracted with ethyl acetate twice. The combined organic layers werewashed with brine, dried over MgSO₄ and concentrated to give yellowishoil. It was purified by Prep. HPLC column to give a colorless oil as TFAsalt (Compound 343). (7 mg, 41% yield)

¹H NMR (CD₃OD, 500 MHz) δ diasteoromer mixture.

LC-MS (retention time: 1.280 min.), MS m/z 678 (MH⁺).

Example 344 Preparation of Compound 344

Step 1: (Scheme 1)

To a solution of 4-p-tolylsulfanylcarbonyl-pyrrolidine-1,2-dicarboxylicacid 1-tert-butyl ester 2-methyl ester (3.0 g, 7.91 mmol) in ethanol (15mL) and THF (30 mL) mixture, sodium borohydride (0.6 g, 15.8 mmol) wasadded. The reaction mixture was stirred at rt. for overnight. Then itwas concentrated, washed with 1 N HCl solution and extracted with EtOActhree times. The organic layers were combined, washed with saturatedNaHCO₃ solution and dried (MgSO₄). Evaporation of solvent gave yellowishoil which was purified by flash column chromatography (silica gel, 3:1EtOAc:Hexanes) to afford colorless oil as product (intermediate 5).(1.77 g, 86% yield)

¹H NMR (CD₃OD, 500 MHz) δ 1.43 (m, 9H), 2.00-2.13 (m, 2H), 2.46 (m, 1H),3.19 (m, 1H), 3.47-3.53 (m, 2H), 3.61 (m, 1H), 3.73 (m, 3H), 4.31 (m,1H).

LC-MS (retention time: 1.240 min.), MS m/z 282 (M+Na⁺).

Step 2: (Scheme 2, step 1)

To a solution of intermediate 5 (80 mg, 0.309 mmol) in THF (10 mL) at 0°C., triphenylphosphine (121.4 mg, 0.463 mmol) and 4-hydroxyquinoline(67.2 mg, 0.463 mmol) were added. Then DEAD (80.6 mg, 0.463 mmol) wasadded. The reaction mixture was warmed to rt. and stirred for 2 days.Then solvent was evaporated and the residue was purified by Prep. HPLCto give colorless oil. It was then dissolved in 4N HCl in dioxane (3 mL)and stirred for 2 hr. Evaporation of solvent gave thick colorless oil asbis HCl salt. (110 mg, 99% yield)

¹H NMR (500 MHz, CD₃OD) δ 2.52 (m, 1H). 2.60 (m, 1H), 3.19 (m, 1H), 3.45(m, 1H), 3.66 (s, 3H), 3.86 (m, 1H), 4.61-4.75 (m, 3H), 7.56 (d, J=6.7Hz, 1H), 7.94 (t, J=7.3 Hz, 1H), 8.10-8.20 (m, 2H), 8.55 (d, J=8.2 Hz,1H), 9.07 (d, J=6.7 Hz, 1H).

LC-MS (retention time: 0.570 min.), MS m/z 287 (MH⁺).

Step 3: (Scheme 2, Step 2)

To a solution of 4-(quinolin-4-yloxymethyl)-pyrrolidine-2-carboxylicacid methyl ester bis hydrochloride salt (110 mg, 0.306 mmol) in CH₃CN(10 mL) was added 2-methoxycarbonylamino-3,3-dimethyl-butyric acid (87mg, 0.46 mmol), DIEA (0.27 mL, 1.53 mmol) and the coupling reagent HOBt(70 mg, 0.46 mmol) and HBTU (174 mg, 0.46 mmol). The solution wasstirred at rt. overnight. Then it was concentrated, washed with waterand extracted with ethyl acetate twice. The combined organic layers werewashed with brine, dried over MgSO₄ and concentrated to give yellowishoil. It was purified by Prep. HPLC column to give colorless oil as TFAsalt. (105 mg, 60% yield)

¹H NMR (CD₃OD, 500 MHz) δ 1.07 (s, 9H). 2.34 (m, 1H), 2.45 (m, 1H), 3.14(m, 1H), 3.27 (s, 3H), 3.75 (s, 3H), 4.05 (m, 1H), 4.20 (m, 1H), 4.31(s, 1H), 4.57-4.63 (m, 2H), 4.73 (m, 1H), 7.53 (d, J=6.7 Hz, 1H), 7.91(t, J=7.6 Hz, 1H), 8.06-8.16 (m, 2H), 8.43 (d, J=8.6 Hz, 1H), 9.02 (d,J=6.4 Hz, 1H).

LC-MS (retention time: 1.250 min.), MS m/z 458 (MH⁺).

Step 4: (Scheme 2, Step 3)

To a solution of1-(2-Methoxycarbonylamino-3,3-dimethyl-butyryl)-4-(quinolin-4-yloxymethyl)-pyrrolidine-2-carboxylicacid methyl ester (100 mg, 0.175 mmol) in THF (6 mL), methanol (3.25 mL)and water (1.0 mL) mixture, lithium hydroxide monohydrate (110 mg, 2.62mmol) was added. The reaction mixture was stirred at rt. for overnight.Then it was acidified with 1N HCl solution to pH=3 to 5 andconcentrated. Extracted with ethyl acetate (3×40 mL) and the organiclayers were combined and dried (MgSO₄). Evaporation of solvent gavethick colorless oil to carry on (25 mg, 32% yield).

To a solution of above compound (25 mg, 0.056 mmol) in CH₃CN (5 mL) wasadded (1R,2S)(1-cyclopropanesulfonyl-aminocarbonyl-2-vinyl-cyclo-propyl)-carbamicacid hydrochloride (22.5 mg, 0.085 mmol), DIEA (0.05 mL, 0.28 mmol) andthe coupling reagent HOBt (12.9 mg, 0.085 mmol) and HBTU (32 mg, 0.085mmol). The solution was stirred at rt. overnight. Then it wasconcentrated, washed with water and extracted with ethyl acetate twice.The combined organic layers were washed with brine, dried over MgSO₄ andconcentrated to give yellow oil. It was purified by Prep. HPLC column togive colorless thick oil as TFA salt (Compound 344). (20 mg, 46% yield)

¹H NMR (CD₃OD, 500 MHz) δ 1.02-1.10 (m, 11H). 1.24 (m, 2H), 1.40 (dd,J=9.2, 5.5 Hz, 1H), 1.90 (dd, J=7.9, 5.5 Hz, 1H), 2.19-2.38 (m, 3H),2.95 (m, 1H), 3.19 (m, 1H), 3.28 (s, 3H), 4.10 (m, 1H), 4.15 (m, 1H),4.34 (s, 1H), 4.55 (m, 1H), 4.62 (d, J=4.6 Hz, 2H), 5.15 (d, J=10.7 Hz,1H), 5.30 (d, J=17.1 Hz, 1H), 5.72 (m, 1H), 7.54 (d, J=6.7 Hz, 1H), 7.93(m, 1H), 8.07-8.18 (m, 2H), 8.41 (d, J=8.6 Hz, 1H), 9.03 (d, J=6.7 Hz,1H), 9.09 (s, 1H).

LC-MS (retention time: 1.617 min.), MS m/z 656 (MH⁺).

Example 345 Preparation of Compound 345

Compound 345 was prepared by following Scheme 2 of Example 344, exceptthat 3-bromophenol was used in the place of 4-hydroxyquinoline in step 1of scheme 2.

Step 1:

To a solution of Intermediate 5 from Example 344 (150 mg, 0.578 mmol) inTHF (15 mL) at 0° C., triphenylphosphine (228 mg, 0.868 mmol) and3-bromophenol (150 mg, 0.868 mmol) were added. Then DEAD (0.14 mL, 0.868mmol) was added. The reaction mixture was warmed to rt. and stirred for2 days. Then solvent was evaporated and the residue was purified byPrep. HPLC to give colorless oil as product. (105 mg, 44% yield)

LC-MS (retention time: 2.023 min.), MS m/z 436 (M+Na⁺).

Step 2:

4-(3-Bromo-phenoxymethyl)-pyrrolidine-1,2-dicarboxylic acid 1-tert-butylester 2-methyl ester (35 mg, 0.085 mmol) was dissolved in 4N HCl indioxane (1.5 mL) and stirred for 2 hr. Evaporation of solvent gave thickcolorless oil. To a solution of this oil in CH₃CN (10 mL) was added2-methoxycarbonylamino-3,3-dimethyl-butyric acid (21.9 mg, 0.1155 mmol),DIEA (0.067 mL, 0.385 mmol) and the coupling reagent HOBt (17.7 mg,0.1155 mmol) and HBTU (43.8 mg, 0.1155 mmol). The solution was stirredat rt. overnight. Then it was concentrated, washed with water andextracted with ethyl acetate twice. The combined organic layers werewashed with brine, dried over MgSO₄ and concentrated. It was thenpurified by Prep. HPLC column to give colorless oil as product. (20 mg,49% yield)

¹H NMR (CD₃OD, 400 MHz) δ 1.03 (s, 9H), 2.15 (m, 1H), 2.24 (m, 1H), 2.83(m, 1H), 3.54 (s, 3H), 3.70 (s, 3H), 3.87 (m, 1H), 3.91-3.98 (m, 3H),4.31 (s, 1H), 4.59 (dd, J=8.80, 5.38 Hz, 1H), 6.89 (d, J=8.32 Hz, 1H),7.03-7.10 (m, 2H), 7.15 (t, J=8.07 Hz, 1H).

LC-MS (retention time: 1.943 min.), MS m/z 485 (MH⁺).

Step 3:

To a solution of4-(3-Bromo-phenoxymethyl)-1-(2-methoxycarbonylamino-3,3-dimethyl-butyryl)-pyrrolidine-2-carboxylicacid methyl ester (17 mg, 0.035 mmol) in THF (1.5 mL), methanol (0.8 mL)and water (0.25 mL) mixture, lithium hydroxide monohydrate (22 mg, 0.525mmol) was added. The reaction mixture was stirred at rt. for 3 days.Then it was acidified with 1N HCl solution to pH=3 to 5 andconcentrated. Extracted with ethyl acetate (2×20 mL) and the organiclayers were combined and dried (MgSO₄). Evaporation of solvent gavethick colorless oil to carry on (15 mg, 91% yield).

To a solution of above acid (15 mg, 0.0318 mmol) in CH₃CN (5 mL) wasadded (1R, 2S)(1-cyclopropanesulfonyl-aminocarbonyl-2-vinyl-cyclo-propyl)-carbamicacid hydrochloride (12.7 mg, 0.0477 mmol), DIEA (0.028 mL, 0.159 mmol)and the coupling reagent HOBt (7.3 mg, 0.0477 mmol) and HBTU (18.1 mg,0.0477 mmol). The solution was stirred at rt. overnight. Then it wasconcentrated, washed with water and extracted with ethyl acetate twice.The combined organic layers were washed with brine, dried over MgSO₄ andconcentrated. It was then purified by Prep. HPLC column to givecolorless thick oil as final product (Compound 345). (14 mg, 64% yield)

¹H NMR (CD₃OD, 400 MHz) δ 1.00-1.06 (m, 11H), 1.21 (m, 2H), 1.37 (dd,J=9.53 Hz, 5.62 Hz, 1H), 1.86 (dd, J=8.07 Hz, 5.62 Hz, 1H), 2.06 (m,1H), 2.14-2.24 (m, 2H), 2.81-2.94 (m, 2H), 3.53 (s, 3H), 3.91-3.97 (m,4H), 4.33 (s, 1H), 4.38 (m, 1H), 5.11 (dd, J=10.27, 1.47 Hz, 1H), 5.28(dd, J=17.12, 1.22 Hz, 1H), 5.70 (m, 1H), 6.89 (m, 1H), 7.05-7.11 (m,2H), 7.16 (t, J=8.07 Hz, 1H).

LC-MS (retention time: 3.500 min.), MS m/z 683 (MH⁺).

Example 346 Preparation of Compound 346

Step 1: (Scheme 1, Step 1)

To a solution of 4-hydroxymethyl-pyrrolidine-1,2-dicarboxylic acid1-tert-butyl ester 2-methyl ester (300 mg, 1.157 mmol) in THF (15 mL) at0° C., triphenylphosphine (455 mg, 1.735 mmol) and 5-bromo-pyridin-3-ol(prepared according to F. E. Ziegler et al., J. Am. Chem. Soc., (1973),95, 7458) (302 mg, 1.735 mmol) were added. Then DEAD (0.273 mL, 1.735mmol) was added. The reaction mixture was warmed to rt. and stirred for2 days. Then solvent was evaporated and the residue was purified byPrep. HPLC to give a yellowish oil. Then it was dissolved in 4N HClsolution in dioxane (3.0 mL) and stirred for 4 hr. Evaporation ofsolvent gave crude product which was further purified by Prep. HPLC toafford a yellowish oil as TFA salt. (70 mg, 11% yield)

LC-MS (retention time: 0.890 min.), MS m/z 315 (MH⁺).

Step 2: (Scheme 1, Step 2)

To a solution of4-(5-Bromo-pyridin-3-yloxymethyl)-pyrrolidine-2-carboxylic acid methylester (70 mg, 0.129 mmol) in CH₃CN (10 mL) was added2-methoxycarbonylamino-3,3-dimethyl-butyric acid (36.5 mg, 0.193 mmol),DIEA (0.135 mL, 0.744 mmol) and the coupling reagent HOBt (30 mg, 0.193mmol) and HBTU (73 mg, 0.193 mmol). The solution was stirred at rt.overnight. Then it was concentrated, washed with water and extractedwith ethyl acetate twice. The combined organic layers were washed withbrine, dried over MgSO₄ and concentrated. It was then purified by Prep.HPLC column to give colorless oil as product. (80 mg, 100% yield)

¹H NMR (CD₃OD, 400 MHz) δ 1.04 (s, 9H), 2.17 (m, 1H), 2.26 (m, 1H), 2.87(m, 1H), 3.50 (s, 3H), 3.70 (s, 3H), 3.88-3.98 (m, 2H), 4.04-4.12 (m,2H), 4.28 (s, 1H), 4.60 (dd, J=9.05, 5.87 Hz, 1H), 7.86 (m, 1H)8.31-8.35 (m, 2H).

LC-MS (retention time: 1.697 min.), MS m/z 486 (MH⁺).

Step 3: (Scheme 1, Step 3)

To a solution of4-(5-Bromo-pyridin-3-yloxymethyl)-1-(2-methoxycarbonylamino-3,3-dimethyl-butyryl)-pyrrolidine-2-carboxylicacid methyl ester (80 mg, 0.133 mmol) in THF (5.6 mL), methanol (3 mL)and water (1 mL) mixture, lithium hydroxide monohydrate (84 mg, 2.0mmol) was added. The reaction mixture was stirred at rt. for 3 days.Then it was acidified with 1N HCl solution to pH=3 to 5. Extracted withethyl acetate (2×20 mL) and the organic layers were combined and dried(MgSO₄). Evaporation of solvent gave thick colorless oil as product(intermediate 6) (50 mg, 80% yield).

¹H NMR (CD₃OD, 400 MHz) δ 1.04 (s, 9H), 2.16-2.30 (m, 2H), 2.88 (m, 1H),3.51 (s, 3H), 3.92 (m, 2H), 4.07 (m, 2H), 4.29 (s, 1H), 4.57 (dd,J=8.56, 5.87 Hz, 1H), 7.79 (m, 1H) 8.29 (m, 2H).

LC-MS (retention time: 1.590 min.), MS m/z 472 (MH⁺).

Step 4: (Scheme 2)

To a solution of4-(5-Bromo-pyridin-3-yloxymethyl)-1-(2-methoxycarbonylamino-3,3-dimethyl-butyryl)-pyrrolidine-2-carboxylicacid (5 mg, 0.0106 mmol) in CH₃CN (5 mL) was added (1R,2S)(1-cyclopropanesulfonyl-aminocarbonyl-2-vinyl-cyclo-propyl)-carbamicacid hydrochloride (4.2 mg, 0.0159 mmol), DIEA (0.009 mL, 0.053 mmol)and the coupling reagent HOBt (2.4 mg, 0.0159 mmol) and HBTU (6.0 mg,0.0159 mmol). The solution was stirred at rt. overnight. Then it wasconcentrated, washed with water and extracted with ethyl acetate twice.The combined organic layers were washed with brine, dried over MgSO₄ andconcentrated. It was then purified by Prep. HPLC column to givecolorless thick oil as final product (Compound 346). (2 mg, 24% yield)

¹H NMR (CD₃OD, 400 MHz) δ 1.00-1.07 (m, 11H), 1.20 (m, 2H), 1.37 (dd,J=9.29 Hz, 5.14 Hz, 1H), 1.86 (dd, J=8.07 Hz, 5.38 Hz, 1H), 2.08 (m,1H), 2.15-2.25 (m, 2H), 2.87-2.94 (m, 2H), 3.51 (s, 3H), 3.92-3.97 (m,2H), 4.02-4.07 (m, 2H), 4.31 (s, 1H), 4.39 (m, 1H), 5.10 (dd, J=10.27,1.47 Hz, 1H), 5.28 (dd, J=17.12, 1.46 Hz, 1H), 5.70 (m, 1H), 7.68 (m,1H), 8.24 (m, 2H).

LC-MS (retention time: 1.727 min.), MS m/z 684 (MH⁺).

Example 347 Preparation of Compound 347

Step 1:

To a solution of intermediate 6 from Example 346 (16 mg, 0.0339 mmol) inDMF (1 mL), 3-thiopheneboronic acid (5.6 mg, 0.044 mmol),tetrakis(triphenylphosphine) palladium (2.0 mg, 0.0017 mmol) and 2MNa₂CO₃ solution (0.051 mL, 0.1017 mmol) were added. The reaction mixturewas heated at 110° C. for 4 hr. Then it was filtered and washed withmethanol. The filtrate was concentrated and purified by Prep.HPLC togive brownish oil as product. (6 mg, 37% yield)

¹H NMR (CD₃OD, 400 MHz) δ 1.05 (s, 9H), 2.21-2.30 (m, 2H), 2.95 (m, 1H),3.42 (s, 3H), 3.93 (m, 1H), 4.01 (m, 1H), 4.20-4.30 (m, 3H), 4.60 (dd,J=8.56, 5.87 Hz, 1H), 7.64 (m, 2H), 8.12 (m, 1H) 8.37 (m, 1H), 8.45 (m,1H), 8.75 (s, 1H).

LC-MS (retention time: 1.353 min.), MS m/z 476 (MH⁺).

Step 2:

To a solution of1-(2-Methoxycarbonylamino-3,3-dimethyl-butyryl)-4-(5-thiophen-3-yl-pyridin-3-yloxymethyl)-pyrrolidine-2-carboxylicacid (6 mg, 0.0126 mmol) in CH₃CN (5 mL) was added (1R,2S)(1-cyclopropanesulfonyl-aminocarbonyl-2-vinyl-cyclo-propyl)-carbamicacid hydrochloride (5.0 mg, 0.0189 mmol), DIEA (0.011 mL, 0.063 mmol)and the coupling reagent HOBt (2.9 mg, 0.0189 mmol) and HBTU (7.2 mg,0.0189 mmol). The solution was stirred at rt. overnight. Then it wasconcentrated, washed with water and extracted with ethyl acetate twice.The combined organic layers were washed with brine, dried over MgSO₄ andconcentrated. It was then purified by Prep. HPLC column to giveyellowish film as TFA salt (Compound 347). (2.2 mg, 22% yield)

¹H NMR (CD₃OD, 400 MHz) δ 1.01-1.07 (m, 11H), 1.19 (m, 2H), 1.36 (m,1H), 1.88 (dd, J=8.07 Hz, 5.62 Hz, 1H), 2.09-2.24 (m, 3H), 2.91 (m, 1H),3.00 (m, 1H), 3.45 (s, 3H), 3.98 (d, J=5.86 Hz, 2H), 4.20-4.31 (m, 3H),4.43 (m, 1H), 5.12 (dd, J=10.27, 1.71 Hz, 1H), 5.29 (dd, J=17.12, 1.22Hz, 1H), 5.69 (m, 1H), 7.64 (m, 2H), 8.11 (m, 1H), 8.31 (m, 1H), 8.43(m, 1H), 8.75 (s, 1H).

LC-MS (retention time: 1.540 min.), MS m/z 688 (MH⁺).

Example 348 Preparation of Compound 348

Step 1: (Scheme 1)

To a solution of intermediate 5 from Example 344 (700 mg, 2.7 mmol) inTHF (90 mL), methanol (50 mL) and water (12 mL) mixture, lithiumhydroxide monohydrate (1700 mg, 2.0 mmol) was added. The reactionmixture was stirred at rt. for overnight. Then it was acidified with 1NHCl solution to pH=3 to 5. Extracted with ethyl acetate (2×20 mL) andthe organic layers were combined and dried (MgSO₄). Evaporation ofsolvent gave thick colorless oil as product (intermediate 7) (0.58, 88%yield).

¹H NMR (CD₃OD, 400 MHz) δ 1.42 (m, 9H), 2.00-2.09 (m, 2H), 2.45 (m, 1H),3.17 (m, 1H), 3.49 (m, 2H), 3.59 (m, 1H), 4.24 (m, 1H).

LC-MS (retention time: 1.08 min.), MS m/z 268 (M+Na⁺).

Step 2: (Scheme 2, Step 1)

To a solution of intermediate 7 (270 mg, 1.1 mmol) in DMSO (10 mL),potassium t-butoxide (309 mg, 2.75 mmol) was added. The reaction mixturewas stirred at rt for 1 hr. Then 2-Bromo-4-chloro-pyridine (254 mg, 1.32mmol) was added. The reaction mixture was stirred at rt for overnight.Then it was quenched with water and washed with ethyl acetate. Theaqueous layer was separated and acidified with 1N HCl solution to pH=3.Extracted with ethyl acetate twice and the organic layers were combinedand dried (MgSO₄). Evaporation of solvent gave an orange oil. It wasthen dissolved in methanol and HCl (gas) was bubbled through for 2 minat −78° C. Then the reaction mixture was warmed to rt and stirred forovernight. Evaporation of solvent gave an orange oil as crude to carryon.

LC-MS (retention time: 0.65 min.), MS m/z 315 (MH⁺).

Step 3: (Scheme 2, step 2)

To a solution of crude4-(2-Bromo-pyridin-4-yloxymethyl)-pyrrolidine-2-carboxylic acid methylester in CH₃CN (20 mL) was added2-methoxycarbonylamino-3,3-dimethyl-butyric acid (312 mg, 1.65 mmol),DIEA (1.15 mL, 6.6 mmol) and the coupling reagent HOBt (252 mg, 1.65mmol) and HBTU (626 mg, 1.65 mmol). The solution was stirred at rt.overnight. Then it was concentrated, washed with water and extractedwith ethyl acetate twice. The combined organic layers were washed withbrine, dried over MgSO₄ and concentrated. It was then purified by Prep.HPLC column to give colorless oil as product. (270 mg, 41% yield twosteps)

¹H NMR (CD₃OD, 400 MHz) δ 1.03 (s, 9H), 2.13-2.19 (m, 2H), 2.87 (m, 1H),3.51 (s, 3H), 3.70 (s, 3H), 3.93 (d, J=6.36 Hz, 2H), 4.11 (m, 2H), 4.27(s, 1H), 4.60 (dd, J=8.80, 5.87 Hz, 1H), 7.06 (d, J=5.87, 2.20 Hz, 1H)7.32 (d, J=2.20 Hz, 1H), 8.18 (d, J=6.11 Hz, 1H).

LC-MS (retention time: 1.657 min.), MS m/z 486 (MH⁺).

Step 4: (Scheme 2, step 3)

To a solution of4-(2-Bromo-pyridin-4-yloxymethyl)-1-(2-methoxycarbonylamino-3,3-dimethyl-butyryl)-pyrrolidine-2-carboxylicacid methyl ester (270 mg, 0.45 mmol) in THF (18 mL), methanol (10 mL)and water (3.3 mL) mixture, lithium hydroxide monohydrate (283 mg, 6.75mmol) was added. The reaction mixture was stirred at rt. for overnight.Then it was concentrated and acidified with 1N HCl solution to pH=3 to5. The off-white solid was collected as product (intermediate 8) (180mg, 85% yield).

¹H NMR (CD₃OD, 500 MHz) δ 1.06 (s, 9H), 2.20-2.29 (m, 2H), 2.89 (m, 1H),3.54 (s, 3H), 3.92 (d, J=6.4 Hz, 2H), 4.06-4.13 (m, 2H), 4.31 (d, J=8.85Hz, 1H), 4.59 (dd, J=8.85, 5.50 Hz, 1H), 7.00 (dd, J=6.10, 2.24 Hz, 1H),7.22 (d, J=1.83 Hz, 1H), 8.12 (d, J=5.80 Hz, 1H).

LC-MS (retention time: 2.113 min.), MS m/z 472 (MH⁺).

Step 5: (Scheme 3)

To a solution of intermediate 8 (10 mg, 0.0212 mmol) in CH₃CN (5 mL) wasadded (1R,2S)(1-cyclopropanesulfonyl-aminocarbonyl-2-vinyl-cyclo-propyl)-carbamicacid hydrochloride (8.5 mg, 0.00318 mmol), DIEA (0.018 mL, 0.106 mmol)and the coupling reagent HOBt (4.9 mg, 0.0318 mmol) and HBTU (12.1 mg,0.0318 mmol). The solution was stirred at rt. overnight. Then it wasconcentrated, washed with water and extracted with ethyl acetate twice.The combined organic layers were washed with brine, dried over MgSO₄ andconcentrated. It was then purified by Prep. HPLC column to givecolorless thick oil as final product (Compound 348). (9 mg, 53% yield)

¹H NMR (CD₃OD, 400 MHz) δ 1.00-1.06 (m, 11H), 1.20 (m, 2H), 1.36 (dd,J=9.54 Hz, 5.38 Hz, 1H), 1.87 (dd, J=8.07 Hz, 5.38 Hz, 1H), 2.04-2.24(m, 3H), 2.88-2.94 (m, 2H), 3.52 (s, 3H), 3.93 (d, J=5.87 Hz, 2H), 4.09(m, 2H), 4.30 (s, 1H), 4.38 (t, J=7.58 Hz, 1H), 5.11 (dd, J=10.27, 1.47Hz, 1H), 5.28 (dd, J=17.12, 1.47 Hz, 1H), 5.70 (m, 1H), 7.00 (dd,J=5.87, 2.20 Hz, 1H), 7.24 (d, J=2.20 Hz, 1H), 8.14 (d, J=5.87 Hz, 1H).

LC-MS (retention time: 1.670 min.), MS m/z 684 (MH⁺).

Example 349 Preparation of Compound 349

Compound 349 was prepared by following scheme of Example 347, exceptthat intermediate 8 from Example 348 was used in the place ofintermediate 6 from Example 346 in step 1.

Step 1:

To a solution of intermediate 8 (20 mg, 0.0423 mmol) in DMF (1 mL),3-thiopheneboronic acid (7.0 mg, 0.055 mmol),tetrakis(triphenylphosphine) palladium (2.4 mg, 0.00212 mmol) and 2MNa₂CO₃ solution (0.063 mL, 0.127 mmol) were added. The reaction mixturewas heated at 110° C. for 30 hr. Then it was filtered and washed withmethanol. The filtrate was concentrated and purified by Prep.HPLC togive brownish oil as product. (10.5 mg, 42% yield) (50177-165)

LC-MS (retention time: 1.690 min.), MS m/z 476 (MH⁺).

Step 2:

To a solution of1-(2-Methoxycarbonylamino-3,3-dimethyl-butyryl)-4-(2-thiophen-3-yl-pyridin-4-yloxymethyl)-pyrrolidine-2-carboxylicacid (10 mg, 0.017 mmol) in CH₃CN (5 mL) was added (1R,2S)(1-cyclopropanesulfonyl-aminocarbonyl-2-vinyl-cyclo-propyl)-carbamicacid hydrochloride (6.8 mg, 0.0254 mmol), DIEA (0.015 mL, 0.085 mmol)and the coupling reagent HOBt (3.9 mg, 0.0254 mmol) and HBTU (9.6 mg,0.0254 mmol). The solution was stirred at rt. overnight. Then it wasconcentrated, washed with water and extracted with ethyl acetate twice.The combined organic layers were washed with brine, dried over MgSO₄ andconcentrated. It was then purified by Prep. HPLC column to give brownishfilm as TFA salt (Compound 349). (2.2 mg, 16% yield). (50177-172)

¹H NMR (CD₃OD, 400 MHz) δ 1.00-1.07 (m, 11H), 1.20 (m, 2H), 1.37 (dd,J=9.04, 5.38 Hz, 1H), 1.88 (dd, J=8.07 Hz, 5.38 Hz, 1H), 2.10-2.25 (m,3H), 2.90 (m, 1H), 3.04 (m, 1H), 3.47 (s, 3H), 3.93-4.02 (m, 2H), 4.29(s, 1H), 4.35-4.45 (m, 3H), 5.12 (d, J=10.51 Hz, 1H), 5.28 (d, J=17.61Hz, 1H), 5.69 (m, 1H), 7.40 (dd, J=6.84, 2.44 Hz, 1H), 7.71-7.80 (m,3H), 8.38 (m, 1H), 8.51 (d, J=7.09 Hz, 1H).

LC-MS (retention time: 1.443 min.), MS m/z 688 (MH⁺).

Example 350 Preparation of Compound 350

Step 1:

To a solution of intermediate 8 from Example 348 (20 mg, 0.0423 mmol) inDMF (2 mL), 2-thiopheneboronic acid (7.0 mg, 0.055 mmol),tetrakis(triphenylphosphine) palladium (2.4 mg, 0.00212 mmol) and bariumhydroxide (40 mg, 0.127 mmol) were added. The reaction mixture washeated at 150° C. in Smith microwave reactor for 110 min. Then it wasfiltered and washed with methanol. The filtrate was concentrated andpurified by Prep.HPLC to give yellowish oil as product. (5.0 mg, 20%yield)

LC-MS (retention time: 2.137 min.), MS m/z 476 (MH⁺).

Step 2:

To a solution of above carboxylic acid (5.0 mg, 0.0085 mmol) in CH₃CN (5mL) was added (1R,2S)(1-cyclopropanesulfonyl-aminocarbonyl-2-vinyl-cyclo-propyl)-carbamicacid hydrochloride (3.4 mg, 0.0127 mmol), DIEA (0.007 mL, 0.0424 mmol)and the coupling reagent HOBt (1.9 mg, 0.0127 mmol) and HBTU (4.8 mg,0.0127 mmol). The solution was stirred at rt. overnight. Then it wasconcentrated, washed with water and extracted with ethyl acetate twice.The combined organic layers were washed with brine, dried over MgSO₄ andconcentrated. It was then purified by Prep. HPLC column to giveyellowish oil as TFA salt (Compound 350). (2.6 mg, 38% yield)

¹H NMR (CD₃OD, 400 MHz) δ 0.99-1.07 (m, 11H), 1.19 (m, 2H), 1.37 (dd,J=9.54, 5.63 Hz, 1H), 1.87 (dd, J=8.07 Hz, 5.38 Hz, 1H), 2.10-2.25 (m,3H), 2.91 (m, 1H), 3.03 (m, 1H), 3.48 (s, 3H), 3.92-4.02 (m, 2H), 4.30(s, 1H), 4.32-4.45 (m, 3H), 5.11 (dd, J=10.27, 1.22 Hz, 1H), 5.28 (d,J=17.11 Hz, 1H), 5.69 (m, 1H), 7.30-7.38 (m, 2H), 7.66 (d, J=2.45 Hz,1H), 7.92 (m, 1H), 7.95 (m, 1H), 8.48 (d, J=6.85 Hz, 1H).

LC-MS (retention time: 2.067 min.), MS m/z 688 (MH⁺).

Example 351 Preparation of Compound 351

Compound 351 was prepared by following scheme of Example 350, exceptthat 3-furanboronic acid was used in the place of 2-thiopheneboronicacid in step 1.

Step 1:

To a solution of intermediate 8 from Example 348 (20 mg, 0.0423 mmol) inDMF (2 mL), 3-furanboronic acid (6.2 mg, 0.055 mmol),tetrakis(triphenylphosphine) palladium (2.4 mg, 0.00212 mmol) and bariumhydroxide (40 mg, 0.127 mmol) were added. The reaction mixture washeated at 150° C. in Smith microwave reactor for 30 min. Then it wasfiltered and washed with methanol. The filtrate was concentrated andpurified by Prep.HPLC to give yellowish oil as product. (12 mg, 49%yield)

LC-MS (retention time: 1.937 min.), MS m/z 460 (MH⁺).

Step 2:

To a solution of above carboxylic acid (5.0 mg, 0.0209 mmol) in CH₃CN (5mL) was added (1R,2S)(1-cyclopropanesulfonyl-aminocarbonyl-2-vinyl-cyclo-propyl)-carbamicacid hydrochloride (8.4 mg, 0.0314 mmol), DIEA (0.018 mL, 0.1046 mmol)and the coupling reagent HOBt (4.8 mg, 0.0314 mmol) and HBTU (11.9 mg,0.0314 mmol). The solution was stirred at rt. overnight. Then it wasconcentrated, washed with water and extracted with ethyl acetate twice.The combined organic layers were washed with brine, dried over MgSO₄ andconcentrated. It was then purified by Prep. HPLC column to giveyellowish oil as TFA salt (Compound 351). (4.0 mg, 24% yield).

¹H NMR (CD₃OD, 400 MHz) δ 1.00-1.08 (m, 11H), 1.21 (m, 2H), 1.37 (dd,J=8.80, 5.62 Hz, 1H), 1.87 (dd, J=8.32 Hz, 5.38 Hz, 1H), 2.11-2.24 (m,3H), 2.91 (m, 1H), 3.03 (m, 1H), 3.49 (s, 3H), 3.91-4.03 (m, 2H), 4.29(s, 1H), 4.35-4.46 (m, 3H), 5.12 (dd, J=10.27, 1.47 Hz, 1H), 5.28 (d,J=17.12 Hz, 1H), 5.69 (m, 1H), 7.11 (m, 1H), 7.38 (dd, J=7.10, 2.69 Hz,1H), 7.71 (d, J=2.69 Hz, 1H), 7.81 (m, 1H), 8.48 (s, 1H), 8.50 (d,J=7.09 Hz, 1H).

LC-MS (retention time: 1.410 min.), MS m/z 672 (MH⁺).

Example 352 Preparation of Compound 352

Compound 352 was prepared by following scheme 2 and scheme 3 of Example348, except that 2,6-dibromopyridine was used in the place of2-bromo-4-chloro-pyridine in step 1 of scheme 2.

Step 1: (Scheme 2, Step 1)

To a solution of intermediate 7 from Example 348 (270 mg, 1.1 mmol) inDMSO (10 mL), potassium t-butoxide (309 mg, 2.75 mmol) was added. Thereaction mixture was stirred at rt for 1 hr. Then 2,6-dibromopyridine(313 mg, 1.32 mmol) was added. The reaction mixture was stirred at rtfor overnight. Then it was quenched with water and washed with ethylacetate. The aqueous layer was separated and acidified with 1N HClsolution to pH=3. Extracted with ethyl acetate twice and the organiclayers were combined and dried (MgSO₄). Evaporation of solvent gave anorange oil. It was then dissolved in methanol and HCl (gas) was bubbledthrough for 2 min at −78° C. Then the reaction mixture was warmed to rtand stirred for overnight. Evaporation of solvent gave an orange oil ascrude to carry on.

LC-MS (retention time: 1.480 min.), MS m/z 315 (MH⁺).

Step 2: (Scheme 2, Step 2)

To a solution of crude4-(6-Bromo-pyridin-2-yloxymethyl)-pyrrolidine-2-carboxylic acid methylester in CH₃CN (20 mL) was added2-methoxycarbonylamino-3,3-dimethyl-butyric acid (312 mg, 1.65 mmol),DIEA (1.15 mL, 6.6 mmol) and the coupling reagent HOBt (252 mg, 1.65mmol) and HBTU (626 mg, 1.65 mmol). The solution was stirred at rt.overnight. Then it was concentrated, washed with water and extractedwith ethyl acetate twice. The combined organic layers were washed withbrine, dried over MgSO₄ and concentrated. It was then purified by flashcolumn chromatography (silica gel, 1:1 hexanes:ethyl acetate) to give acolorless oil as product. (340 mg, 63% yield two steps)

¹H NMR (CD₃OD, 400 MHz) δ 1.03 (s, 9H), 2.10-2.24 (m, 2H), 2.84 (m, 1H),3.55 (s, 3H), 3.70 (s, 3H), 3.83 (m, 1H), 3.94 (m, 1H), 4.20-4.29 (m,2H), 4.31 (s, 1H), 4.59 (dd, J=8.80, 5.13 Hz, 1H), 6.76 (d, J=8.07 Hz,1H) 7.11 (d, J=7.58 Hz, 1H), 7.53 (t, J=7.83 Hz, 1H).

LC-MS (retention time: 1.820 min.), MS m/z 486 (MH⁺).

Step 3: (Scheme 2, Step 3)

To a solution of4-(6-Bromo-pyridin-2-yloxymethyl)-1-(2-methoxycarbonylamino-3,3-dimethyl-butyryl)-pyrrolidine-2-carboxylicacid methyl ester (330 mg, 0.679 mmol) in THF (28 mL), methanol (15 mL)and water (5 mL) mixture, lithium hydroxide monohydrate (427 mg, 10.18mmol) was added. The reaction mixture was stirred at rt. for 2 days.Then it was concentrated and acidified with 1N HCl solution to pH=3 to5. The white solid was collected as product (intermediate 9) (310 mg,97% yield).

¹H NMR (CD₃OD, 500 MHz) δ 1.06 (s, 9H), 2.18-2.25 (m, 2H), 2.88 (m, 1H),3.57 (s, 3H), 3.84 (m, 1H), 3.96 (m, 1H), 4.25 (m, 1H), 4.28-4.35 (m,2H), 4.58 (m, 1H), 6.79 (d, J=7.94 Hz, 1H), 7.13 (d, J=7.32 Hz, 1H),7.55 (m, 1H).

LC-MS (retention time: 3.030 min.), MS m/z 472 (MH⁺).

Step 4: (Scheme 3)

To a solution of intermediate 9 (10 mg, 0.0212 mmol) in CH₃CN (5 mL) wasadded (1R,2S)(1-cyclopropanesulfonyl-aminocarbonyl-2-vinyl-cyclo-propyl)-carbamicacid hydrochloride (8.5 mg, 0.00318 mmol), DIEA (0.018 mL, 0.106 mmol)and the coupling reagent HOBt (4.9 mg, 0.0318 mmol) and HBTU (12.1 mg,0.0318 mmol). The solution was stirred at rt. overnight. Then it wasconcentrated, washed with water and extracted with ethyl acetate twice.The combined organic layers were washed with brine, dried over MgSO₄ andconcentrated. It was then purified by Prep. HPLC column to giveyellowish film as TFA salt (Compound 352). (10.2 mg, 60% yield).

¹H NMR (CD₃OD, 400 MHz) δ 1.00-1.06 (m, 11H), 1.20 (m, 2H), 1.37 (dd,J=9.54 Hz, 5.63 Hz, 1H), 1.86 (dd, J=8.07 Hz, 5.38 Hz, 1H), 2.05 (m,1H), 2.12-2.25 (m, 2H), 2.86-2.94 (m, 2H), 3.54 (s, 3H), 3.87 (m, 1H),3.94 (m, 1H), 4.18-4.27 (m, 2H), 4.33 (s, 1H), 4.37 (m, 1H), 5.11 (dd,J=10.27, 1.72 Hz, 1H), 5.28 (dd, J=17.12, 1.47 Hz, 1H), 5.70 (m, 1H),6.76 (d, J=8.32 Hz, 1H), 7.11 (d, J=7.33 Hz, 1H), 7.53 (t, J=7.82 Hz,1H).

LC-MS (retention time: 1.837 min.), MS m/z 684 (MH⁺).

Example 353 Preparation of Compound 353

Compound 353 was prepared by following scheme of Example 347, exceptthat intermediate 9 from Example 352 was used in the place ofintermediate 6 from Example 346 in step 1.

Step 1:

To a solution of intermediate 9 (25 mg, 0.053 mmol) in DMF (1 mL),3-thiopheneboronic acid (8.8 mg, 0.0688 mmol),tetrakis(triphenylphosphine) palladium (3.1 mg, 0.00265 mmol) and 2MNa₂CO₃ solution (0.080 mL, 0.159 mmol) were added. The reaction mixturewas heated at 110° C. for overnight. Then it was filtered and washedwith methanol. The filtrate was concentrated and purified by Prep.HPLCto give brownish oil as product. (15 mg, 48% yield)

¹H NMR (CD₃OD, 500 MHz) δ 1.06 (s, 9H), 2.20-2.31 (m, 2H), 2.94 (m, 1H),3.55 (s, 3H), 3.91 (m, 1H), 3.98 (m, 1H), 4.34 (s, 1H), 4.37-4.46 (m,2H), 4.61 (dd, J=8.85, 5.19 Hz, 1H), 6.77 (d, J=8.24 Hz, 1H), 7.39 (d,J=7.32 Hz, 1H), 7.48 (dd, J=5.19, 3.05 Hz, 1H), 7.68 (dd, J=4.88, 1.22Hz, 1H), 7.77 (t, J=7.93 Hz, 1H), 8.04 (m, 1H).

LC-MS (retention time: 1.857 min.), MS m/z 476 (MH⁺).

Step 2:

To a solution of1-(2-Methoxycarbonylamino-3,3-dimethyl-butyryl)-4-(6-thiophen-3-yl-pyridin-2-yloxymethyl)-pyrrolidine-2-carboxylicacid (15 mg, 0.0254 mmol) in CH₃CN (5 mL) was added (1R,2S)(1-cyclopropanesulfonyl-aminocarbonyl-2-vinyl-cyclo-propyl)-carbamicacid hydrochloride (10.2 mg, 0.0382 mmol), DIEA (0.022 mL, 0.127 mmol)and the coupling reagent HOBt (5.8 mg, 0.0382 mmol) and HBTU (14.5 mg,0.0382 mmol). The solution was stirred at rt. overnight. Then it wasconcentrated, washed with water and extracted with ethyl acetate twice.The combined organic layers were washed with brine, dried over MgSO₄ andconcentrated. It was then purified by Prep. HPLC column to giveyellowish film as TFA salt (Compound 353). (6 mg, 29% yield)

¹H NMR (CD₃OD, 400 MHz) δ 1.00-1.06 (m, 11H), 1.20 (m, 2H), 1.36 (dd,J=9.29, 5.38 Hz, 1H), 1.86 (dd, J=8.07 Hz, 5.38 Hz, 1H), 2.07 (m, 1H),2.16-2.25 (m, 2H), 2.87-2.99 (m, 2H), 3.54 (s, 3H), 3.87-3.99 (m, 2H),4.31-4.44 (m, 4H), 5.11 (dd, J=10.27, 1.46 Hz, 1H), 5.28 (d, J=17.12 Hz,1H), 5.70 (m, 1H), 6.67 (d, J=8.31 Hz, 1H), 7.33 (d, J=7.34 Hz, 1H),7.44 (dd, J=4.89, 2.93 Hz, 1H), 7.63-7.70 (m, 2H), 7.99 (m, 1H).

LC-MS (retention time: 2.770 min.), MS m/z 688 (MH⁺).

Example 354 Preparation of Compound 354

Compound 354 was prepared by following scheme of Example 347, exceptthat intermediate 9 from Example 352 was used in the place ofintermediate 6 from Example 346 and phenyl boronic acid was used in theplace of 3-thiopheneboronic acid in step 1.

Step 1:

To a solution of intermediate 9 (20 mg, 0.0423 mmol) in DMF (1 mL),phenyl boronic acid (6.7 mg, 0.0688 mmol), tetrakis(triphenylphosphine)palladium (2.4 mg, 0.00212 mmol) and Cs₂CO₃ (41 mg, 0.127 mmol) wereadded. The reaction mixture was heated at 110° C. for overnight. Then itwas filtered and washed with methanol. The filtrate was concentrated andpurified by Prep.HPLC to give yellowish oil as product. (12 mg, 49%yield)

LC-MS (retention time: 2.733 min.), MS m/z 470 (MH⁺).

Step 2:

To a solution of1-(2-Methoxycarbonylamino-3,3-dimethyl-butyryl)-4-(6-phenyl-pyridin-2-yloxymethyl)-pyrrolidine-2-carboxylicacid (12 mg, 0.0206 mmol) in CH₃CN (5 mL) was added (1R,2S)(1-cyclopropanesulfonyl-aminocarbonyl-2-vinyl-cyclo-propyl)-carbamicacid hydrochloride (8.2 mg, 0.0308 mmol), DIEA (0.018 mL, 0.1028 mmol)and the coupling reagent HOBt (4.7 mg, 0.0308 mmol) and HBTU (11.7 mg,0.0308 mmol). The solution was stirred at rt. overnight. Then it wasconcentrated, washed with water and extracted with ethyl acetate twice.The combined organic layers were washed with brine, dried over MgSO₄ andconcentrated. It was then purified by Prep. HPLC column to give a whitesolid as TFA salt (Compound 354). (1.5 mg, 9% yield)

¹H NMR (CD₃OD, 400 MHz) δ 1.00-1.07 (m, 11H), 1.20 (m, 2H), 1.36 (m,1H), 1.85 (m, 1H), 2.09 (m, 1H), 2.17-2.25 (m, 2H), 2.87-3.00 (m, 2H),3.52 (s, 3H), 3.84-4.00 (m, 2H), 4.33-4.44 (m, 4H), 5.11 (dd, J=10.27,1.71 Hz, 1H), 5.28 (d, J=17.12, 1.22 Hz, 1H), 5.70 (m, 1H), 6.71 (d,J=8.31 Hz, 1H), 6.78 (m, 1H), 7.34-7.44 (m, 3H), 7.74 (m, 1H), 7.95 (m,1H), 8.01 (d, J=8.31 Hz, 1H).

LC-MS (retention time: 3.553 min.), MS m/z 682 (MH⁺).

Example 355 Preparation of Compound 355

Compound 354 was prepared by following scheme of Example 347, exceptthat intermediate 9 from Example 352 was used in the place ofintermediate 6 from Example 346 and 3-furan boronic acid was used in theplace of 3-thiopheneboronic acid in step 1.

Step 1:

To a solution of intermediate 9 (20 mg, 0.0423 mmol) in DMF (1 mL),3-furan boronic acid (6.2 mg, 0.055 mmol), tetrakis(triphenylphosphine)palladium (2.4 mg, 0.002115 mmol) and 2M Na₂CO₃ solution (0.064 mL,0.127 mmol) were added. The reaction mixture was heated at 110° C. for 2days. Then it was filtered and washed with methanol. The filtrate wasconcentrated and purified by Prep.HPLC to give yellowish oil as product.(7.0 mg, 29% yield)

Step 2:

To a solution of above carboxylic acid (6.0 mg, 0.0109 mmol) in CH₃CN (5mL) was added (1R,2S)(1-cyclopropanesulfonyl-aminocarbonyl-2-vinyl-cyclo-propyl)-carbamicacid hydrochloride (4.4 mg, 0.0163 mmol), DIEA (0.0095 mL, 0.0544 mmol)and the coupling reagent HOBt (2.5 mg, 0.0163 mmol) and HBTU (6.2 mg,0.0163 mmol). The solution was stirred at rt. overnight. Then it wasconcentrated, washed with water and extracted with ethyl acetate twice.The combined organic layers were washed with brine, dried over MgSO₄ andconcentrated. It was then purified by Prep. HPLC column to giveyellowish film as TFA salt (Compound 355). (1.5 mg, 18% yield)

¹H NMR (CD₃OD, 400 MHz) δ 0.98-1.07 (m, 11H), 1.20 (m, 2H), 1.35 (dd,J=9.54, 5.87 Hz, 1H), 1.86 (dd, J=8.07 Hz, 5.62 Hz, 1H), 2.06 (m, 1H),2.15-2.25 (m, 2H), 2.85-2.98 (m, 2H), 3.55 (s, 3H), 3.89 (m, 1H), 3.95(m, 1H), 4.28-4.42 (m, 4H), 5.11 (dd, J=10.27, 1.71 Hz, 1H), 5.28 (dd,J=17.12, 1.22 Hz, 1H), 5.69 (m, 1H), 6.63 (d, J=8.07 Hz, 1H), 6.90 (m,1H), 7.16 (d, J=7.33 Hz, 1H), 7.53 (m, 1H), 7.63 (m, 1H), 8.08 (s, 1H).

LC-MS (retention time: 3.340 min.), MS m/z 672 (MH⁺).

Example 356 Preparation of Compound 356

Compound 356 was prepared by following scheme of Example 350, exceptintermediate 9 from Example 352 was used in the place of intermediate 8from Example 348 in step 1.

Step 1:

To a solution of intermediate 9 (20 mg, 0.0423 mmol) in DMF (2 mL),2-thiopheneboronic acid (7.0 mg, 0.055 mmol),tetrakis(triphenylphosphine) palladium (2.4 mg, 0.00212 mmol) and bariumhydroxide (40 mg, 0.127 mmol) were added. The reaction mixture washeated at 150° C. in Smith microwave reactor for 30 min. Then it wasfiltered and washed with methanol. The filtrate was concentrated andpurified by Prep.HPLC to give brownish oil as product. (13.0 mg, 52%yield)

¹H NMR (CD₃OD, 400 MHz) δ 1.03 (s, 9H), 2.18-2.25 (m, 2H), 2.93 (m, 1H),3.55 (s, 3H), 3.83 (m, 1H), 3.98 (m, 1H), 4.34 (s, 1H), 4.38 (m, 2H),4.58 (dd, J=8.05, 5.14 Hz, 1H), 6.63 (d, J=8.07 Hz, 1H), 7.07 (dd,J=4.89, 3.67 Hz, 1H), 7.33 (d, J=7.34 Hz, 1H), 7.42 (d, J=5.14 Hz, 1H),7.60-7.66 (m, 2H).

LC-MS (retention time: 3.393 min.), MS m/z 476 (MH⁺).

Step 2:

To a solution of1-(2-Methoxycarbonylamino-3,3-dimethyl-butyryl)-4-(6-thiophen-2-yl-pyridin-2-yloxymethyl)-pyrrolidine-2-carboxylicacid (11.5 mg, 0.0195 mmol) in CH₃CN (5 mL) was added (1R,2S)(1-cyclopropanesulfonyl-aminocarbonyl-2-vinyl-cyclo-propyl)-carbamicacid hydrochloride (7.8 mg, 0.0293 mmol), DIEA (0.017 mL, 0.0975 mmol)and the coupling reagent HOBt (4.5 mg, 0.0293 mmol) and HBTU (11.1 mg,0.0293 mmol). The solution was stirred at rt. overnight. Then it wasconcentrated, washed with water and extracted with ethyl acetate twice.The combined organic layers were washed with brine, dried over MgSO₄ andconcentrated. It was then purified by Prep. HPLC column to give anoff-white solid as TFA salt (Compound 356). (8.5 mg, 54% yield)

¹H NMR (CD₃OD, 400 MHz) δ 0.99-1.06 (m, 11H), 1.21 (m, 2H), 1.36 (dd,J=9.54, 5.38 Hz, 1H), 1.86 (dd, J=8.07 Hz, 5.62 Hz, 1H), 2.06 (m, 1H),2.15-2.25 (m, 2H), 2.87-2.99 (m, 2H), 3.54 (s, 3H), 3.89 (m, 1H), 3.96(m, 1H), 4.30-4.44 (m, 4H), 5.11 (dd, J=10.51, 1.71 Hz, 1H), 5.28 (dd,J=17.11, 1.22 Hz, 1H), 5.70 (m, 1H), 6.62 (d, J=8.07 Hz, 1H), 7.07 (dd,J=4.89, 3.66 Hz, 1H), 7.33 (d, J=7.59 Hz, 1H), 7.42 (d, J=4.89 Hz, 1H),7.60-7.66 (m, 2H).

LC-MS (retention time: 1.967 min.), MS m/z 688 (MH⁺).

Example 357 Preparation of Compound 357

Step 1:

To a solution of (2S,4R) Fmoc-4-amino-1-boc-pyrrolidine-2-carboxylicacid (400 mg, 0.884 mmol) in acetonitrile (15 mL), five drops ofpyrrolidine was added. The reaction mixture was stirred at rt for 3 hr.Then it was concentrated and put on high vacuum to give crude4-amino-1-boc-pyrrolidine-2-carboxylic acid. In another round-bottomedflask, a solution of Pd₂dba₃ (40 mg, 5% mol) and racemic-BINAP (56 mg,10% mol) was stirred under nitrogen in degassed toluene (8 mL) at rt for1 h. Then 1-chloroisoquinoline (216 mg, 1.326 mmol) and sodiumt-butoxide (340 mg, 3.536 mmol) were added and the reaction mixture wasstirred for 30 min. Then 4-amino-1-boc-pyrrolidine-2-carboxylic acid wasadded and the reaction mixture was heated under reflux for 1 h. Waterwas added to quench the reaction and the aqueous layer was separated andfiltered through filter paper. It was then concentrated and purified byPrep. HPLC to give coupled product as TFA salt. (165 mg, 40% yield)

¹H NMR (CD₃OD, 400 MHz) δ 1.44 (m, 9H), 2.51-2.74 (m, 2H), 3.64 (m, 1H),4.01 (m, 1H), 4.49 (m, 1H), 4.64 (m, 1H), 7.30 (d, J=6.85 Hz, 1H), 7.58(d, J=6.85 Hz, 1H), 7.79 (m, 1H), 7.91-7.99 (m, 2H), 8.56 (d, J=8.56 Hz,1H).

LC-MS (retention time: 1.707 min.), MS m/z 358 (MH⁺).

Step 2:

To a solution of 4-(Isoquinolin-1-ylamino)-pyrrolidine-1,2-dicarboxylicacid 1-tert-butyl ester (115 mg, 0.244 mmol) in CH₃CN (10 mL) was added(1R,2S)(1-cyclopropanesulfonyl-aminocarbonyl-2-vinyl-cyclo-propyl)-carbamicacid hydrochloride (97 mg, 0.366 mmol), DIEA (0.255 mL, 1.464 mmol) andthe coupling reagent HOBt (56 mg, 0.366 mmol) and HBTU (139 mg, 0.366mmol). The solution was stirred at rt. overnight. Then it wasconcentrated, washed with water and extracted with ethyl acetate twice.The combined organic layers were washed with brine, dried over MgSO₄ andconcentrated to give yellow oil. It was purified by Prep. HPLC column toa yellowish solid as TFA salt (112 mg, 67% yield).

¹H NMR (CD₃OD, 400 MHz) δ 1.05 (m, 2H), 1.20 (m, 2H), 1.40-1.48 (m,10H), 1.87 (dd, J=8.19, 5.50 Hz, 1H), 2.23 (m, 1H), 2.39 (m, 1H), 2.50(m, 1H), 2.93 (m, 1H), 3.65 (m, 1H), 4.08 (m, 1H), 4.33 (t, J=7.09 Hz,1H), 4.69 (m, 1H), 5.12 (d, J=10.27 Hz, 1H), 5.29 (d, J=17.12 Hz, 1H),5.74 (m, 1H), 7.31 (d, J=6.85 Hz, 1H), 7.60 (d, J=7.09 Hz, 1H), 7.80 (m,1H), 7.93-8.00 (m, 2H), 8.56 (d, J=8.19 Hz, 1H).

LC-MS (retention time: 2.023 min.), MS m/z 570 (MH⁺).

Step 3:

2-(1-Cyclopropanesulfonylaminocarbonyl-2-vinylcyclopropyl-carbamoyl)-4-(isoquinolin-1-ylamino)-pyrrolidine-1-carboxylicacid tert-butyl ester (31 mg, 0.0453 mmol) was dissolved in 4N HCl indioxane (1.5 mL) and stirred at rt. for 2 hr. Evaporation of solventgave yellowish oil as bis hydrochloride salt. To a solution of bishydrochloride salt in CH₃CN (5 mL) was added N-boc-L-t-leucine (11.5 mg,0.0498 mmol), DIEA (0.047 mL, 0.272 mmol) and the coupling reagent HOBt(10.4 mg, 0.068 mmol) and HBTU (25.8 mg, 0.068 mmol). The solution wasstirred at rt. overnight. Then it was concentrated, washed with waterand extracted with ethyl acetate twice. The combined organic layers werewashed with brine, dried over MgSO₄ and concentrated to give yellowishoil. It was purified by Prep. HPLC column to give an off-white solid asfinal product (Compound 357). (9 mg, 29% yield)

¹H NMR (CD₃OD, 400 MHz) δ 0.98 (m, 2H), 1.05 (s, 9H), 1.20 (m, 2H),1.36-1.43 (m, 10H), 1.84 (m, 1H), 2.10-2.30 (m, 2H), 2.52 (m, 1H), 2.90(m, 1H), 4.07 (m, 1H), 4.17-4.27 (m, 2H), 4.47 (m, 1H), 4.79 (m, 1H),5.07 (d, J=9.29 Hz, 1H), 5.24 (d, J=16.87 Hz, 1H), 5.72 (m, 1H), 6.62(m, 1H), 6.98 (d, J=6.11 Hz, 1H), 7.47 (m, 1H), 7.62 (m, 1H), 7.69 (d,J=8.07 Hz, 1H), 7.84 (d, J=5.87 Hz, 1H), 8.20 (d, J=8.56 Hz, 1H).

LC-MS (retention time: 2.043 min.), MS m/z 683 (MH⁺).

Section H LC-MS Condition for Section H

Columns:

-   -   (Method A)—YMC ODS S7 C18 3.0×50 mm    -   (Method B)—YMC ODS-A S7 C18 3.0×50 mm    -   (Method C)—YMC S7 C18 3.0×50 mm    -   (Method D)—YMC Xterra ODS S7 3.0×50 mm    -   (Method E)—YMC Xterra ODS S7 3.0×50 mm    -   (Method F)—YMC ODS-A S7 C18 3.0×50 mm    -   (Method H)—Xterra S7 3.0×50 mm    -   (Method I)—Xterra S7 C18 3.0×50 mm    -   (Method G)—YMC C18 S5 4.6×50 mm    -   (Method J)—Xterra ODS S7 3.0×50 mm    -   (Method K)—YMC ODS-A S7 C18 3.0×50 mm

Gradient: 100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B

Gradient time: 2 min. (A, B, D, F, G, H, I); 8 min. (C, E); 4 min (J); 3min (K)

Hold time: 1 min. (A, B, D, F, G, H, I, J, K); 2 min. (C, E)

Flow rate: 5 mL/min (A, B, C, D, E, F, G)

Flow rate: 4 mL/min (J, K)

Detector Wavelength: 220 nm

Solvent A: 10% MeOH/90% H₂O/0.1% TFA

Solvent B: 10% H₂O/90% MeOH/0.1% TFA.

Example 370 Preparation of Compound 370

Step 1:

A solution of (1R,2S)/(1S,2R)-1-amino-2-vinylcyclopropane carboxylicacid ethyl ester hydrochloride (2.54 g, 12 mmol) in CH₃CN (70 mL) wastreated with a solution of diisopropylethylamine (9.5 mL, 67 mmol),[(4R)-(2-methoxycarbonyl-7-methoxylquinoline-4-oxo)-S-proline](5.9 g,13.2 mmol), and TBTU (3.89 g, 12.21 mmol) in CH₃CN (50 mL). The reactionmixture was stirred for 14 h and concentrated. The residue dissolved inEtOAc was repeatedly washed with NaHCO₃ (aq.), brine, dried (MgSO₄), andconcentrated. The residue was purified over Biotage 65M column(EtOAc/hexane: 45-100%) to provide the high Rf stereo isomer(Boc-P2[(4R)-(2-methoxycarbonyl-7-methoxylquinoline-4-oxo)-S-proline]-P1(1R,2SVinyl Acca) Acid ethyl ester 2.0 g (52%) as a white solid: ¹H NMR(methanol-d₄) δ ppm 1.24 (t, J=7.02 Hz, 3H), 1.38 (m, 11H), 1.76 (m,1H), 2.21 (m, 1H), 2.45 (m, 1H), 2.71 (m, 1H), 3.92 (m, 2H), 3.96 (s,3H), 4.03 (s, 3H), 4.16 (q, J=7.22 Hz, 2H), 4.42 (m, 1H), 5.10 (m, 1H),5.30 (m, 1H), 5.44 (s, 1H), 5.77 (m, 1H), 7.27 (d, J=9.16 Hz, 1H), 7.48(s, 1H), 7.52 (s, 1H), 8.05 (s, 1H).

Step 2:

A solution of the high Rf product (3.16 g, 5.40 mmol) of Step 1 ofExample 370{Boc-P2[(4R)-(2-methoxycarbonyl-7-methoxylquinoline-4-oxo)-S-proline]-P1(1R,2SVinyl Acca) COOEt} at 0° C. dissolved in MeOH/THF (1/1, 13.2 mL) wastreated with aqueous 1.0 N NaOH (5.5 mL, 5.5 mmol), stirred for 1 h,neutralized by the addition of AcOH. The solvent was removed in vacuo.The residue was redissolved in THF/CH₂Cl₂ (1/1, 150 mL), dried (MgSO₄)and concentrated in vacuo to provide the product which was directly usedin next step: LC-MS (retention time: 1.53 Method D), MS m/z 570 (M⁺+1).

Step 3:

To a solution of the product (assumed at 5.4 mmol) of step 2 example 370at 0° C. dissolved in THF (35 mL) was added a solution of fresh madeCH₂N₂ (30 mmol) in Et₂O (80 mL). The reaction mixture was stirred at thetemperature for 0.5 h, and stirred at rt for 18.5 h. After bubblingnitrogen for 1 h to the reaction mixture, the solution was removed invacuo. The residue redissolved in EtOAc (1 L) was washed with saturatedNaHCO₃ (aq.), (2×200 mL), brine (100 mL), and dried (MgSO₄). The solventwas removed in vacuo to afford the product 3.10 g (97% two steps): LC-MS(retention time: 3.06, Method J), MS m/z 594 (M⁺+1).

Step 4:

To a solution of the product (3.03 g, 5.10 mmol) of step 3 of example370{Boc-P2[(4R)-(2-diazoacetyl-7-methoxylquinoline-4-oxo)-S-proline]-P1(1R,2SVinyl Acca) COOEt} at 0° C. dissolved in THF (110 mL) was added 2 mL of48% HBr. The mixture was stirred for 1 h, partitioned between EtOAc (500mL) and saturated NaHCO₃ (aq.) (100 mL). The EtOAC layer was separated,dried (MgSO₄). The solvent was removed to afford the product (3.12 g,95%): LC-MS (retention time: 1.56 Method D). MS m/z 648 (M⁺+1), MS m/z646 (M⁻−1).

Step 5:

The product (1.0 g, 1.54 mmol) of step 4 of example 370{Boc-P2[(4R)-(2-bromoacetyl-7-methoxylquinoline-4-oxo)-S-proline]-P1(1R,2SVinyl Acca) COOEt} was treated with isopropylthiourea (0.365 g, 3.09mmol) in isopropyl alcohol (57 mL) for 2 h, and then the solvent wasremoved. The residue dissolved in aqueous 1.0 N HCl (30 mL) and EtOAC(200 mL) was adjusted pH to 7 by addition of 1.0 N NaOH (aq.). Theaqueous layer was extracted with EtOAc (2×100 mL) and the combinedextract was dried (MgSO₄), concentrated. The residue was purified byover Biotage 40+M column (EtOAc-hexanes: 30-100%) to afford the product870 mg (84%) and ready for the next step.

Step 6:

The product (0.250 g, 0.375 mmol) of step 5 of example 370 {Boc-P2{(4R)-[2-(2-isopropylaminothiazol-4-yl)-7-methoxylquinoline-4-oxo]-S-proline}-P1(1R,2SVinyl Acca) COOEt} was treated with 4N HCl/dioxane (2.5 mL, 10 mmol) for2.5 h and concentrated in vacuo. To the residue was addedN-methylmorpholine (0.206 mL, 1.875 mmol) in DMF (3 mL),N-Boc-L-tert-leucine (0.117 g, 0.506 mmol), and HATU (0.192 g, 0.506mmol). The mixture was stirred overnite and partitioned between EtOAcand pH 4.0 buffer. The EtOAc layer was washed with water, NaHCO₃ (aq.),dried (MgSO₄), concentrated. The residue was purified over a Biotage 40Mcolumn (MeOH—CH₂Cl₂: 0-8%) to afford the product 0.289 g (99%):

LC-MS (retention time: 2.53, Method K), MS m/z 779 (M⁺+1).

Step 7:

To a suspension of the product of Step 6 (274 mg, 0.352 mmol) of Example370{BOCNH-P3(L-t-BuGly)-{[2-(2-isopropylaminothiazol-4-yl)-7-methoxylquinoline-4-oxo]-S-proline}-P1(1R,2SVinyl Acca)-COOEt} in THF (10.6 mL), CH₃OH (2.6 mL), and H₂O (5.3 mL)was added LiOH (0.068 g, 2.86 mmol). The reaction mixture was stirredfor 24, adjusted to pH 6, removed the organic solvents in vacuo. Theaqueous residue was acidified to pH 4, and extracted with CH₂Cl₂repeatedly. Combined organic solvent was dried (MgSO₄), and concentratedin vacuo to afford the desired product 255 mg (95%): LC-MS (retentiontime: 2.58, Method K), MS m/z 751 (M⁺+1).

Step 8:

A solution of CDI (0.024 g, 0.15 mmol) and the product of Step 7 ofExample 370 (0.0683 g, 0.09 mmol){BOCNH-P3(L-t-BuGly)-{[2-(2-isopropylaminothiazol-4-yl)-7-methoxylquinoline-4-oxo]-S-proline}-P1(1R,2SVinyl Acca)-COOH} in THF (2 mL) was refluxed for 60 min and allowed tocool down to rt. Cyclopropanesulfonamide (0.022 g. 0.18 mmol) was addedfollowed by the addition of neat DBU (0.027 mL, 0.18 mmol). The reactionwas stirred for overnite, worked up by diluting with EtOAc and washedwith pH 4.0 buffer, dried (MgSO₄), and concentrated. The residue waspurified repeatedly by preparative HPLC (0-100% solvent B) and over 1000μM preparative TLC plate from Analtech (20×40 cM) to afford 0.0032 g(4%) the desired product (Compound 370) as a pale yellow foam:

LC-MS (Retention time: 1.71, method I) MS m/z 854 (M⁺+1).

Example 371 Preparation of Compound 371

Step 1:

To a suspension of N-Boc-cis-L-4-Hydroxyproline methyl ester (10 g, 40.7mmol) and 7-chloroquinolin-4-ol (8.73 g, 49.0 mmol) in THF (200 mL)cooled to 0° C. was added PPh₃ (12.82 g, 48.9 mmol) and DIAD. (8.80 g,42.13 mmol). The mixture was slowly allowed to warm to rt overnite,stirred at total of 30 h. The mixture was dissolved in EtOAc (800 mL),washed with 1N aqueous HCl, 5% aqueous K₂CO₃ (3×100 mL), brine (2×100mL) and dried (MgSO₄), and concentrated. The residue was purifiedseveral times over a Biotage 65M (MeOH—CH₂Cl₂: 0-10%) to affordcumulatively 10.57 g (68%) of the desired product as a glass: ¹H NMR(CDCl₃) δ 1.40 (s, 9H), 2.33-2.42 (m, 1H), 2.61-2.72 (m, 1H), 3.75 (s,3H), 3.91 (m, 2H), 4.45-4.59 (m, 1H), 5.13 (m, 1H), 6.61-6.64 (m, 1H),7.41 (dd, J=9, 2 Hz, 1H), 7.98 (d, J=2 Hz, 1H), 8.03 (d, J=9 Hz, 1H),8.67-8.71 (m, 1H). LC-MS (retention time: 1.39, method D), MS m/e 407(M⁺+1).

Step 2:

To a solution of the product (10.57 g, 26.0 mmol) of Step 1 of Example371 {BOC—N-P2[(4R)-(7-chloroquinoline-4-oxo) proline methyl ester}dissolved in MeOH (800 mL) cooled to 0° C. was added an aqueous 1N NaOHsolution (44.5 mL, 44.5 mmol). The mixture was warmed to rt after 6 h,stirred overnite, and the pH adjusted to pH 7 using 1.0 N aqueous HCl.The solution was concentrated until only the water layer remained, thepH adjusted to 4 using 6N aqueous HCl and the mixture was partitionedrepeatedly with EtOAc (3×500 mL). The combined organic layers were dried(MgSO₄) and concentrated to afford 10.16 g (100%) of the as a whitesolid. ¹H NMR (DMSO-d₆) δ 1.32, 1.34 (two s (rotamers) 9H), 2.31-2.40(m, 1H), 2.58-2.69 (m, 1H), 3.65-3.81 (m, 2H), 4.33-4.40 (m, 1H), 5.34(m, 1H) 7.10-7.11 (m, 1H), 7.57 (d, J=9 Hz, 1H), 7.98 (s, 1H), 8.09-8.14(m, 1H), 8.75 (d, J=5 Hz, 1H), 12.88 (brs, 1H). ¹³C NMR (DMSO-d₆) δ27.82, 35.84, 51.52, 57.75, 76.03, 79.33, 102.95, 119.54, 123.86,126.34, 127.24, 134.49, 149.32, 152.88, 153.25, 159.08, 173.74. LC-MS(retention time: 1.48, method D), MS m/e 393 (M⁺+1)

Step 3:

To a solution of the product (5.11 g, 13 mmol) of Step 2 of Example 371{Boc-4(R)-(7-chloroquinoline-4-oxo) proline}, the HCl salt (3.48 g, 18.2mmol) of vinyl Acca (existing as a 1:1 mixture of diastereoisomers(1R,2S/1S,2R where cyclopropyl carboxyethyl group is syn to vinylmoiety) and NMM (7.1 mL 65 mmol) in DMF (30 mL) was added HATU (6.92 g,18.2 mmol). The mixture was stirred for 3 days. The reaction mixture wasdiluted with EtOAc (180 mL) and was partitioned with pH 4.0 buffer(3×100 mL). The organic layer was washed with saturated aqueous NaHCO₃(2×50 mL), water (2×50 mL), and brine (2×50 mL). The organic solutionwas dried (MgSO₄) and concentrated. The residue was purified over aBiotage 40M column (EtOAc-Hexanes: 50% to 100%) to afford 2.88 g of theproduct existing as a diastereomeric mixture. This mixture was partiallyseparated using a Biotage 65M column (MeOH-EtOAc: 0% to 9%) to affordBOC—NH—P2[(4R)-(7-chloroquinoline-4-oxo)-S-proline]-P1(1R,2S vinyl accaP1 moiety)-COOEt as the initial eluted high Rf isomer (1.20 g, 17.4%).¹H NMR (CDCl₃/Methanol-d₄) δ 1.16 (t, J=7 Hz, 3H), 1.35 (s, 9H),1.37-1.43 (m, 1H), 1.76-1.84 (m, 1H), 2.06-2.11 (m, 1H), 2.35-2.45 (m,1H), 2.63 (m, 1H), 3.72-3.93 (m, 2H), 4.02-4.15 (m, 1H), 4.33-4.40 (m,1H), 5.06 (d, J=9 Hz, 1H), 5.16 (m, 1H), 5.24 (d, J=17 Hz, 1H),5.63-5.70 (m, 1H), 6.74 (m, 1H), 7.39 (dd, J=9, 2 Hz, 1H), 7.74-7.78 (m,1H), 7.89 (d, J=2 Hz, 1H), 7.97 (d, J=9 Hz, 1H), 8.60 (d, J=5 Hz, 1H).¹H NMR (methanol-d₄, 60/40 Rotomers) δ 1.24 (t, J=7 Hz, 3H), 1.39, 1.43(2s, 9H, ratio 4:6), 1.71-1.74 (m, 0.4H), 178-1.81 (m, 0.6H), 2.18-2.23(m, 1H), 2.65-2.69 (m, 0.4H), 2.71-2.76 (m, 0.6H), 3.88-3.96 (m, 2H),4.11-4.18 (m, 2H), 4.39-4.45 (m, 1H), 5.09-5.13 (m, 1H), 5.28-5.33 (m,1H), 5.37 (m, 1H), 5.73-5.81 (m, 1H), 7.05 (d, J=5 Hz, 1H), 7.53 (d,J=8.9 Hz, 1H), 7.92 (s, 1H), 8.12 (d, J=8.9 Hz, 1H), 8.70 (d, J=5 Hz,1H). LC-MS (retention time: 1.54, method A) MS m/z 530 (M⁺+1). The restof the material (˜1.66 g, 24%) was mixed fractions greatly enriched inthe lower Rf isomer.

Step 4:

The product (0.65 g, 1.22 mmol) of step 3 of Example 371{BOC-P2[(4R)-(7-chloroquinoline-4-oxo)-S-proline]-P1(1R,2S VinylAcca-CO₂Et} was dissolved in 4N HCl/dioxane (4.5 ml, 18 mmol) andstirred for 1 h at rt. The reaction mixture was concentrated and thecrude product was directly used in next step: LC-MS (retention time:0.94, method A) LC-MS m/z 430 (M⁺+1).

Step 5:

To suspension of the product 1(0.22 mmol) of step 4 Example 371 {Bis HClSalt of NH₂—P2[(4R)-(7-chloroquinoline-4-oxo)-S-proline]-P1(1R,2S-VinylAcca)-COOEt), N—BOC-L-tert-leucine (BOC L-tBuGly) (0.34 g, 1.47 mmol),DIPEA (1.0 ml, 5.74 mmol), HOBT.H₂O (0.22 g, 1.47 mmol) in CH₂Cl₂ (15mL) was added HBTU (0.56 g, 1.47 mmol) at rt. The reaction mixture wasstirred overnite, diluted with CH₂Cl₂ (50 mL), washed with pH 4.0 buffer(2×20 mL), saturated aqueous NaHCO₃ (50 mL), brine (50 mL), dried(MgSO₄), and concentrated. The residue was purified over a Biotage 40 Mcolumn (EtOAc-Hexanes: 15% to 60%) to afford 607 mg (77%) of the productas a foam. ¹H NMR (CDCl₃-methanol-d₄) δ 1.00 (s, 9H), 1.19 (t, J=7 Hz,1H), 1.30 (s, 9H), 1.38 (m, 1H), 1.78-1.83 (m, 1H), 2.01-2.46 (m, 2H),2.73-2.82 (m 1H), 3.96-4.03 (m, 1H), 4.04 (d, J=10 Hz, 1H), 4.11 (q, J=7Hz, 2H), 4.42 (d, J=12 Hz, 1H), 4.68-4.73 (m, 1H), 5.09-5.13 (m, 1H),5.23-5.31 (m, 2H), 5.67-5.79 (m, 1H), 6.78 (d, J=9 Hz, 1H), 7.38 (d, J=9Hz, 1H), 7.70 (s, 1H), 7.96 (s, 1H), 8.08 (d, J=9 Hz, 1H), 8.68 (d, J=5Hz, 1H). LC-MS (retention time: 1.64, method A), MS m/z 643 (M⁺+1).

Step 6:

To a suspension of the product (207 mg, 0.32 mmol) of Step 5 of Example371{BOCNH-P3(L-t-BuGly)-P2[(4R)-7-chloroquinoline-4-oxo)-S-proline]-P1(1R,2SVinyl Acca)-CO₂Et} in THF (14 mL), CH₃OH (2 mL), and H₂O (8 mL) wasadded LiOH (62 mg, 2.60 mmol). The reaction mixture was stirred for oneday, adjusted to neutral pH, and concentrated in vacuo until only theaqueous layer remained. The resulting aqueous residue was acidified topH 4.0 by addition of 1.0 N aqueous HCl and then saturated with solidNaCl. This aqueous mixture was extracted repeatedly with EtOAc (3×60mL), the combined organic solvent was dried (Mg₂SO₄) and concentrated invacuo to afford 107 mg (54%) of the product{BOCNH-P3(L-t-BuGly)-P2[(4R)-(7-chloroquinoline-4-oxo)-S-proline]-P1(1R,2SVinyl Acca)-CO₂H} as a white solid. ¹H NMR (CDCl₃) δ 1.06 (s, 9H), 1.23(2s, 9H), 1.31-1.43 (m 1H), 1.63-1.70 (m, 1H), 1.85-1.89 (m, 1H), 2.19(m, 1H), 2.65-2.78 (m, 1H), 4.03-4.10 (m, 1H), 4.18-4.21 (m, 1H),4.55-4.62 (m, 1H), 5.03-5.12 (m, 1H), 5.23-5.31 (m, 1H), 5.51 (m, 1H),5.88-5.95 (m, 1H), 7.12 (m, 1H), 7.47-7.50 (m, 1H), 7.96 (m, 1H), 8.26(d, J=9 Hz, 1H), 8.75 (d, J=5 Hz, 1H). LC-MS (retention time: 1.46,method A), MS m/z 615 (M⁺+1).

Step 7:

To a solution of the tripeptide acid (0.0453 g, 0.074 mmol) of Step 6Example 371{BOCNH-P3(L-t-BuGly)-P2[(4R)-(7-chloroquinoline-4-oxo)-S-proline]-P1(1R,2SVinyl Acca)-CO₂H} in THF (4 mL) was added CDI (17 mg, 0.10 mmol), andthe resulting solution refluxed for 45 min and allowed to cool down tort. Cyclopropylsulfonamide (0.013 g, 0.10 mmol) was added in one portionbefore the addition of neat DBU (0.015 mL, 0.10 mmol). The reaction wasstirred for 18 h, diluted with EtOAc (200 mL) and washed pH 4.0 buffer(3×30 mL), water (2×30 mL), brine (30 mL), dried (MgSO₄) and purifiedusing one 20×40 cM 1000 Analtech PTLC plate (MeOH—CH₂Cl₂: 0 to 2%) toafford the desired product (Compound 371) as a foam (0.040 g, 76%): ¹HNMR δ 0.95-1.23 (m, 4H), 1.03 (s, 9H), 1.19 (s, 9H), 1.40-1.43 (m, 1H),1.85 (dd, J=8, 5 Hz, 1H), 2.12-2.20 (m, 1H), 2.43 (m, 1H), 2.82 (m, 1H),4.07-4.19 (m, 2H), 4.51-4.57 (m, 2H), 5.07 (d, J=10 Hz, 1H), 5.25 (d,J=17 Hz, 1H), 5.85 (m, 1H), 5.48 (s, 1H), 7.09 (d, J=5 Hz, 1H), 7.45 (d,J=9 Hz, 1H), 7.92 (m, 1H), 8.20 (d, J=9 Hz, 1H), 8.72 (d, J=5 Hz, 1H);LC-MS (retention time: 1.52, method B), MS m/z 718 (M⁺+1). HRMS m/z(M+H)⁺ calcd for C₄₁H₅₁N₅SO₉: 718.2677 found 718.2674.

Example 372 Preparation of Compound 372

Step 1:

To a solution of L-tert-leucine (2 g, 15.25 mmol) dissolved in CH₃CN (50mL) was added TMSCN (7.06 mL, 56.41 mmol) and stirred for 15 min. Thereaction mixture was heated to 75° C. for 30 min. Cyclopentylchloroformate (2.83 g, 19.06 mmol) was added to the reaction mixture andthe reaction mixture was heated at 80° C. overnite, concentrated invacuo. The residue was treated with MeOH (40 mL), stirred for 10 min,and concentrated in vacuo. The residue was adjusted pH to 8.5, andextracted with Et₂O (2×200 mL). The aqueous layer was acidified to pH 3and extracted with CH₂Cl₂ (2×200 mL). The combined extract was dried(MgSO₄), and concentrated in vacuo. The residue was recrystallized fromminimal amount of Et₂O/hexanes to afford the product 3.48 g (94%): ¹HNMR (500 MHz, methanol-d₄) δ ppm 1.00 (s, 9H), 1.59 (m, 2H), 1.73 (m,4H), 1.84 (dd, J=5.95, 3.20 Hz, 2H), 3.98 (s, 1H), 5.02 (m, 1H).

Step 2:

To a solution of the product (530.1 mg, 1.04 mmol) of Step 4 of Example371 {HCl salt of P2[(4R)-7-chloroquinoline-4-oxo)-S-proline]-P1(1R,2SVinyl Acca) COOEt, the product (328 mg, 1.35 mmol) of Step 1 of Example372 {(L)-2-cyclopentyloxycarbonylamino-3,3-dimethyl-butyric acid}, HOBT(146 mg, 1.08 mmol), and diisopropylethylamine (0.755 mL, 4.32 mmol) inCH₂Cl₂ (7 mL) was added HBTU (512 mg, 1.35 mmol). The reaction mixturewas stirred for overnite and partitioned between CH₂Cl₂ and pH 4.0buffer. The CH₂Cl₂ layer was washed with water, saturated NaHCO₃ (aq.),dried (MgSO₄), concentrated. The residue was purified over a Biotage 40Mcolumn (EtOAc-Hexanes: 35-100%) to afford the product 640 mg (92%): ¹HNMR (methanol-d₄) δ ppm 1.02 (s, 9H), 1.26 (m, 4H), 1.56 (m, 10H), 2.19(q, J=8.75 Hz, 1H), 2.41 (m, 1H), 2.70 (dd, J=14.19, 8.09 Hz, 1H), 4.01(dd, J=11.90, 3.05 Hz, 1H), 4.13 (m, 2H), 4.20 (s, 1H), 4.53 (m, 1H),4.62 (m, 1H), 5.09 (d, J=10.38 Hz, 1H), 5.26 (d, J=17.09 Hz, 1H), 5.47(m, 1H), 5.77 (m, 1H), 7.07 (d, J=5.49 Hz, 1H), 7.47 (m, 1H), 7.94 (m,1H), 8.20 (d, J=8.85 Hz, 1H), 8.72 (d, J=5.49 Hz, 1H). LC-MS (retentiontime: 1.71, Method B), MS m/z 655 (M⁺+1).

Step 3:

Tripeptide acid was prepared by following Step 7 of Scheme 2 of Example370, except that cyclopentoxycarbonyl—NH-P3(L-tert-BuGly)-P2[(4R)-(7-chloroquinoline-4-oxo)-S-proline]-P1(1R,2SVinyl Acca)-COOEt used in place of the product of Step 6 of Example 370.

Modification: 0.636 g (0.97 mmol) of the product of Step 2 of Example372 used, 0.424 g obtained (69% yield).

Product:

Data: ¹H NMR (methanol-d₄) δ ppm 1.02 (s, 9H), 1.57 (m, 11H), 2.14 (q,J=9.03 Hz, 1H), 2.46 (m, 1H), 2.68 (m, 1H), 4.02 (dd, J=11.89, 3.11 Hz,1H), 4.19 (m, 1H), 4.50 (d, J=26.35 Hz, 1H), 4.64 (t, J=8.42 Hz, 1H),5.04 (m, 1H), 5.24 (d, J=17.20 Hz, 1H), 5.44 (s, 1H), 5.87 (m, 1H), 7.05(d, J=5.12 Hz, 1H), 7.48 (m, 1H), 7.92 (m, 1H), 8.18 (d, J=8.78 Hz, 1H),8.71 (d, J=5.49 Hz, 1H). LC-MS (retention time: 2.32, Method A), MS m/z627 (M⁺+1).

Step 4:

A solution of CDI (0.021 g, 0.13 mmol) and the product of Step 3 ofExample 372 (0.058 g, 0.09 mmol){BOCNH-P3(L-t-BuGly)-P2[(4R)-7-chloroquinoline-4-oxo)-S-proline]-P1(1R,2SVinyl Acca)-CO₂H} in THF (2 mL) was refluxed for 40 min and allowed tocool down to rt. A total of 0.016 g (0.13 mmol) ofcyclopropanesulfonamide, followed by the addition of a solution of neatDBU (0.019 mL, 0.13 mmol). The reaction was stirred for overnite, thendiluted with EtOAc (100 mL) and washed pH 4.0 buffer (2×), dried(MgSO₄), concentrated and purified over three 1000 μM preparative TLCplate from Analtech (20×40 cM, eluted sequentially with 50% to 0% to 2%MeOH in CH₂Cl₂) to provide product (Compound 372) 27.3 mg (40%): ¹H NMR(methanol-d₄) δ ppm 0.94 (m, 2H), 1.02 (s, 9H), 1.14 (m, 1H), 1.49 (m,11H), 1.86 (m, 1H), 2.14 (m, 1H), 2.49 (m, 1H), 2.68 (dd, J=13.89, 7.48Hz, 1H), 2.78 (m, 1H), 4.08 (m, 1H), 4.22 (s, 1H), 4.55 (m, 2H), 5.05(d, J=10.07 Hz, 1H), 5.22 (d, J=17.09 Hz, 1H), 5.46 (m, 1H), 5.86 (m,1H), 7.07 (d, J=5.19 Hz, 1H), 7.46 (d, J=8.55 Hz, 1H), 7.91 (s, 1H),8.18 (d, J=8.85 Hz, 1H), 8.72 (d, J=5.19 Hz, 1H). LC-MS (retention time:1.52 Method I), MS m/z 730 (M⁺+1).

Example 373 Preparation of Compound 373

Step 1:

A solution of 2-amino-4-methoxylacetophenone (4.45 g, 26.94 mmol) at 0°C. dissolved in CH₂Cl₂ (100 mL) was treated with cyclopropanecarbonylchloride (3.1 mL, 33.68 mmol) diisopropylethylamine (19 mL, 107.8 mmol),DMAP (0.780 g, 6.4 mmol). The reaction mixture was stirred at rtovernite and concentrated in vacuo. The residue dissolved in CH₂Cl₂ (500mL) was washed with aqueous 1 N HCl, water, NaHCO₃ (aq.), and dried(MgSO₄). The solvent was removed in vacuo and the solid residue wastreated with EtOAc/hexanes (1/1) to provide the product (5.35 g, 85%):

¹H NMR (methanol-d₄) δ ppm 0.94 (m, 4H), 1.69 (m, J=3.97 Hz, 1H), 2.60(s, 3H), 3.84 (s, 3H), 6.69 (d, J=7.93 Hz, 1H), 7.98 (d, J=8.85 Hz, 1H),8.23 (s, 1H).

Step 2:

A solution of product (5.35 g, 22.72 mmol) of Step 1 example 373{cyclopropanecarboxylic acid (2-acetyl-5-methoxy-phenyl)-amide} andtert-BuOK (5.45 g, 48.6 mmol) in tert-butanol (130 g) was refluxed for 6h. The reaction mixture was cooled, poured into ice cold buffer andadjusted to pH 7, filtered. The solid collection was recrystallized fromMeOH/Et₂O to provide the product (1 g, 20%): ¹H NMR (methanol-d₄) δ ppm0.96 (m, 2H), 1.15 (m, 2H), 1.94 (m, 1H), 3.87 (s, 3H), 5.86 (m, 1H),6.93 (m, 2H), 8.04 (d, J=8.85 Hz, 1H).

Step 3:

To a solution of N-Boc-L-3-hydroxyproline (1.06 g, 4.32 mmol) andtriphenylphosphine (2.27 g, 8.64 mmol) at 0° C. dissolved THF (25 mL)was added a solution of the product (0.93 g, 4.32 mmol) of Step 2Example 373 {2-Cyclopropyl-7-methoxy-quinolin-4-ol} and DEAD (1.50 g,8.64 mmol) in THF (25 mL) over 30 min. The reaction mixture was stirredovernite and concentrated. The residue was purified twice by a Biotage40+M column (EtOAc-Hexanes: 20-65%) to afford the product 1.74 g (90%):LC-MS (retention time: 2.56, Method J), MS m/z 443 (M⁺+1).

Step 4:

To a suspension of (1.70 g, 3.86 mmol) of the product of Step 3 ofExample 373(Boc-(4R)-(2-cyclopropyl-7-methoxy-quinoline-4-oxo)-S-proline methylester} in THF (91 mL), CH₃OH (18.2 mL), and H₂O (27 mL) was added LiOH(0.73 g, 30 mmol). The reaction mixture was stirred for 16 h, adjustedto pH 6, the organic solvent was removed in vacuo. The residue wasacidified to pH 4, and extracted with EtOAc (4×100 mL). The combinedorganic extract was dried (MgSO₄), and concentrated in vacuo to supplythe product 1.64 g (100%): ¹H NMR (methanol-d₄) δ ppm 1.32 (m, 13H),2.37 (m, 2H), 2.71 (m, 1H), 3.86 (m, 1H), 3.95 (s, 3H), 4.14 (m, 1H),4.43 (m, 1H), 5.41 (s, 1H), 6.65 (s, 1H), 7.19 (m, 1H), 7.30 (m, 1H),8.02 (dd, J=12.63, 9.33 Hz, 1H).

Step 5:

The product (1.61 g, 2.79 mmol) of Step 4 of Example 373{Boc-P2{(4R)-[2-cyclopropyl-7-methoxylquinoline-4-oxo]-S-proline}-P1(1R,2SVinyl Acca) COOEt} was dissolved in HCl/dioxane (15 mL; 60 mmol) andstirred for 3 h at rt. The reaction mixture was concentrated andazeotroped with dry THF to afford the product (1.58 g, 100%): LC-MS(retention time: 2.12, Method K), MS m/z 566 (M⁺+1).

Step 6:

To a suspension of the product (1.58 g, 2.79 mmol) of Step 5 of Example373 {Bis HCl salt of P2{(4R)-[2-cyclopropyl-7-methoxylquinoline-4-oxo]-S-proline}-P1(1R,2SVinyl Acca) COOEt}, diisopropylethylamine (1.65 mL, 9.25 mmol),N-Boc-L-tert-leucine (0.775 g, 3.35 mmol), HOBT.H₂O (0.515 g, 3.36 mmol)in CH₂Cl₂ (13 mL) was added HBTU (1.28 g, 3.36 mmol). The mixture wasstirred for 14 h and partitioned between EtOAc and pH 4.0 buffer. TheEtOAc layer was dried (MgSO₄), concentrated. The residue was purifiedover a Biotage 40+M column (EtOAc-hexanes: 20-100%, followed MeOH) andfurther purified by 20×40 cM 1000 Analtech PTLC plate (MeOH—CH₂Cl₂ 2%)to afford the product 1.4 g (63%):

¹H NMR (methanol-d₄) δ ppm 1.04 (s, 9H), 1.20 (m, 5H), 1.28 (s, 9H),1.39 (m, 2H), 1.69 (m, 1H), 2.19 (m, 2H), 2.36 (m, 1H), 2.63 (dd,J=13.54, 7.68 Hz, 1H), 3.90 (s, 3H), 4.08 (m, 4H), 4.19 (d, J=11.34 Hz,1H), 4.47 (d, J=11.71 Hz, 1H), 4.56 (t, J=8.60 Hz, 1H), 5.08 (m, 1H),5.24 (m, 1H), 5.39 (s, 1H), 5.78 (m, 1H), 6.56 (s, 1H), 6.96 (dd,J=9.15, 2.20 Hz, 1H), 7.21 (d, J=2.56 Hz, 1H), 7.97 (d, J=9.15 Hz, 1H).LC-MS (retention time: 2.34, Method K), MS m/z 679 (M⁺+1).

Step 7:

To a suspension of the product of Step 6 of Example 373 (1.28 g, 1.89mmol),Boc-NH-P3(L-tert-BuGly)-P2[(4R)-(2-cyclopropyl-7-methoxylquinoline-4-oxo)-S-proline]-P1(1R,2SVinyl Acca)-COOEt, in THF (93 mL), CH₃OH (23 mL), and H₂O (45 mL) wasadded LiOH (0.491 g, 20.4 mmol). The reaction mixture was stirred for18.5 h, adjusted to pH 4, removed the organic solvent in vacuo. Theresidue was extracted with EtOAc (5×100 mL). Combined organic solventwas dried (MgSO₄), and concentrated in vacuo to afford the desiredproduct 1.17 g (97%): ¹H NMR (methanol-d₄) δ ppm 1.04 (s, 9H), 1.24 (s,9H), 1.27 (m, 3H), 1.42 (m, 2H), 1.68 (dd, J=8.05, 5.12 Hz, 1H), 2.17(m, 1H), 2.33 (m, 1H), 2.47 (m, 1H), 2.66 (m, 1H), 3.95 (s, 3H), 4.09(m, 2H), 4.51 (d, J=11.71 Hz, 1H), 4.59 (t, J=8.60 Hz, 1H), 5.07 (m,1H), 5.26 (m, 1H), 5.52 (s, 1H), 5.85 (m, 1H), 6.69 (s, 1H), 7.10 (dd,J=9.15, 2.20 Hz, 1H), 7.27 (d, J=2.20 Hz, 1H), 8.10 (d, J=9.15 Hz, 1H).LC-MS (retention time: 2.21, Method K), MS m/z 651 (M⁺+1).

Step 8:

A solution of CDI (0.058 g, 0.344 mmol) and the product of Step 7 ofExample 373 (0.160 g, 0.246 mmol){Boc-NH-P3(L-tert-BuGly)-P2[(4R)-(2-cyclopropyl-7-methoxylquinoline-4-oxo)-S-proline]-P1(1R,2SVinyl Acca)-COOH} in THF (2 mL) was refluxed for 60 min and allowed tocool down to rt. Cyclopropanesulfonamide (0.041 g, 0.344 mmol) followedby the addition of neat DBU (0.051 mL, 0.344 mmol). The reaction wasstirred for 24 h and worked up by partition the reaction mixture betweenpH 4.0 buffer and EtOAc. The organic layer was dried (MgSO₄),concentrated and purified by preparative HPLC (0-100% solvent B) tosupply the product (Compound 373) 0.086 g (46%): ¹H NMR (TRIFLUOROACETICACID-D) δ ppm 1.04 (s, 9H), 1.21 (m, 16H), 1.41 (m, 1H), 1.87 (dd,J=8.05, 5.49 Hz, 1H), 2.26 (m, 3H), 2.61 (dd, J=12.99, 6.77 Hz, 1H),2.93 (m, 1H), 3.92 (s, 3H), 4.09 (m, 1H), 4.21 (m, 1H), 4.49 (m, 2H),5.11 (d, J=11.71 Hz, 1H), 5.27 (d, J=17.20 Hz, 1H), 5.46 (s, 1H), 5.76(m, 1H), 6.62 (m, 2H), 7.01 (dd, J=8.97, 2.01 Hz, 1H), 7.23 (d, J=2.56Hz, 1H), 8.00 (d, J=8.78 Hz, 1H).

Example 374 Preparation of Compound 374

To a solution of m-anisidine (58 g, 471 mmol) in 800 mL of CH₃CN wasadded Meldrum's acid (75 g, 518 mmol), and trimethylformate (60 g, 565mmol). The heterogeneous mixture was refluxed for 2 h. The solvent wasremoved in vacuo, MeOH (30 mL) was added, and the resulting precipitatewas filtered and washed with 10-15 mL of MeOH. The MeOHaddition/filtration procedure was repeated on the concentrated motherliquor. The resulting combined solid was dried (˜20 torr, 45° C.overnite) to afford 117.6 g (90%) of the intermediate5-[(3-Methoxyphenyl-amino)methylene]-2,2-dimethyl-[1,3]dioxane-4,6-dione.

Step 2:

To a solution of Ph₂O (500 g) heated to 250° C. was added 108.7 g (392mmol) of5-[(3-Methoxyphenyl-amino)methylene]-2,2-dimethyl-[1,3]dioxane-4,6-dionein portions over a 30 min period. The mixture was heated an additional15 min, cooled to rt, diluted with hexanes (800 mL) and the resultingslurry stirred overnite. The hexanes was decanted off, the solid residuedissolved in 600 mL of MeOH at reflux, cooled to rt and the resultingsolid filtered and washed with minimal CH₂Cl₂. The analogousrecrystallization procedure was followed to afford a total of 20.73 g(30%) of 7-methoxyquinolin-4-ol as a light brown solid. ¹H NMR(methanol-d₄) δ 3.87 (s, 3H), 6.23 d, J=7.3 Hz, 1H), 6.68 (d, J=2.4 Hz,1H), 6.96 (dd, J=9.0, 2.4 Hz, 1H), 8.11 (d, J=9 Hz, 1H); LC-MS(retention time: 0.77, method D), MS m/z 176 (M⁺+1).

Step 3:

To a solution of) of N-Boc-cis-L-4-Hydroxyproline methyl ester (12.24 g,49.8 mmol) and PPh₃ (26.14 g, 99.7 mmol) in THF (200 mL) cooled to 0° C.was added a solution of DEAD (17.36 g, 99.7 mmol) and7-methoxyquinolin-4-ol (8.73 g, 49.8 mmol) in (THF 700 mL) over a 45 minperiod. The mixture was slowly allowed to warm to rt overnite,concentrated in vacuo. The residue was purified over a Biotage 65Mcolumn (MeOH-EtOAc: 0-10%) to afford 12.78 g (64%) of the product as acolorless glass: ¹H NMR (CDCl₃) δ 1.36 (s, 9H), 2.26-2.35 (m, 1H),2.57-2.68 (m, 1H), 3.71 (s, 3H), 3.75-3.92 (m, 2H), 3.86, 3.87 (two s(rotamers) 3H), 4.41-4.53 (m, 1H), 5.09 (m, 1H), 6.52 (d, J=5.5 Hz, 1H),7.06-7.09 (m, 1H), 7.24-7.26 (m, 1H), 7.94 (d, J=9.1 Hz, 1H), 8.50-8.56(m, 1H); LC-MS (retention time: 1.34, method D),

MS m/e 403 (M+11).

Step 4:

To a solution of the product (8.54 g, 21.2 mmol) of step 3 of Example374 {BOC—N—P2[(4R)-(7-methoxyquinoline-4-oxo) proline methyl ester} in600 mL of 5:1 THF/MeOH was added a solution of LiOH (4.0 g, 167 mmol) in150 mL of water. The mixture was stirred overnite, the pH was adjustedto pH 7 using 6N aqueous HCl, and the solution concentrated until onlythe water layer remained. The residue was adjusted to pH 4 using 1Naqueous HCl, NaCl added to saturate the mixture and was partitionedrepeatedly with first EtOAc and then THF as the product was aqueoussoluble. The combined organic layers were dried (MgSO₄) and concentratedto afford the product 8.18 g (99%) as a white solid. ¹H NMR(CDCl₃-Methanol-d₄) δ 1.42 (s, 9H), 2.40-2.49 (m, 1H), 2.68-2.77 (m,1H), 3.88 (m, 2H), 3.94 (s, 3H), 4.41-4.53 (m, 1H), 5.32 (m, 1H),6.86-6.92 (m, 1H), 7.21 (dd, J=9, 2 Hz, 1H), 7.30 (d, J=2 Hz, 1H),8.05-8.10 (m, 1H), 8.62 (d, J=6 Hz, 1H); LC-MS (retention time 1.20,method A), MS m/z 389 (M⁺+1).

Step 5:

To a solution of the product (4.50 g, 11.60 mmol) of Step 4 of Example374 {Boc-4(R)-(7-methoxyquinoline-4-oxo) proline}, 2.66 g (13.9 mmol) ofthe HCl salt of vinyl Acca (existing as a 1:1 mixture ofdiastereoisomers (1R,2S/1S,2R where cyclopropyl carboxyethyl group issyn to vinyl moiety), 10 mL (57.4 mmol) of DIPEA, and 2.13 g (13.9 mmol)of HOBT.H₂O in 150 mL of CH₂Cl₂ was added 5.27 g (13.9 mmol) of HBTU,and the mixture stirred overnite. The solution was diluted with 200 mLof CH₂Cl₂ and was partitioned with pH 4.0 buffer (2×50 mL). The organiclayer was washed with saturated aqueous NaHCO₃ (2×50 mL), water (2×50mL), and brine (2×50 mL). The organic solution was dried (MgSO₄),concentrated and purified using a Biotage 65M column (eluted with 0-9%MeOH/EtOAc) to provide ofBOC—NH-P2[(4R)-(7-methoxyquinoline-4-oxo)-S-proline]-P1(1R,2S vinyl accaP1 moiety)-COOEt as the initial eluted isomer (2.21 g, 36% overall),followed by 1.13 g (19%) of pure lower Rf isomerBOC—NH-P2[(4R)-(7-methoxyquinoline-4-oxo)-S-proline]-P1(1S,2R Vinyl AccaP1 moiety)-CO₂Et. Mixed fractions were also obtained. Data forBOCN-P2[(4R)-(7-methoxyquinoline-4-oxo)-S-proline]-P1(1R,2S)-(VinylAcca)-COOEt:¹H NMR (CDCl₃) δ 1.16 (t, J=7 Hz, 3H), 1.35 (s, 9H), 1.37-1.47 (m, 1H),1.74-1.88 (m, 1H), 2.04-2.13 (m, 1H), 2.32-2.46 (m, 1H), 2.58-2.69 (m,1H), 3.76 (m, 1H), 3.87 (s, 3H), 4.02-4.13 (m, 2H), 4.30-4.44 (m, 1H),5.05-5.19 (m, 2H), 5.24 (d, J=17 Hz, 1H), 5.63-5.71 (m, 1H), 6.61 (m,1H), 7.07 (dd, J=9, 2 Hz, 1H), 7.22 (d, J=2 Hz, 1H), 7.76-7.83 (m, 1H),7.92 (d, J=9 Hz, 1H), 8.50 (d, J=5 Hz, 1H). LC-MS (retention time: 1.38,method A), MS m/z 526 (M⁺+1).

Step 6:

A total of product (1.35 g, 2.90 mmol) of Step 5 of Example 374{BOC-P2[(4R)-(7-methoxyquinoline-4-oxo)-S-proline]-P1(1R,2S VinylAcca)-COOEt} was dissolved in 4N HCl/dioxane (15 ml, 60 mmol) and wasstirred for 2.5 h at rt. The reaction mixture was concentrated in vacuoto supply 1.3 g (100%) of the product as a tan solid which was directlyused in next step. ¹H NMR (methanol-d₄) δ 1.25 (t, J=7 Hz, 1H),1.47-1.52 (m, 1H), 1.78 (dd, J=8, 5 Hz, 1H), 2.21-2.32 (m, 1H),2.55-2.64 (m, 1H), 2.99 (dd, J=15, 7 Hz, 1H), 3.96 (s, 2H), 4.06 (s,3H), 4.14 (q, J=7 Hz, 2H), 4.69-4.75 (m, 1H), 5.13 (d, J=10 Hz, 1H),5.33 (d, J=17 Hz, 1H), 5.71-5.83 (m, 1H), 5.89 (m, 1H), 7.44 (m, 1H),7.49-7.52 (m, 1H), 8.51-8.55 (m, 1H), 8.94-8.96 (m, 1H); ¹³C NMR(methanol-d₄) δ 14.62, 23.08, 30.89, 34.73, 36.97, 41.03, 52.42, 57.11,60.17, 62.70, 81.13, 100.06, 103.07, 117.02, 118.53, 122.70, 126.86,134.74, 143.15, 146.75, 166.62, 167.71, 169.37, 171.18. LC-MS (retentiontime: 0.94, method D), MS m/z 426 (M+1)

Step 7:

To suspension of product (1.3 g, 2.61 mmol) of Step 6 of Example 374{NH₂—P2[(4R)-(7-methoxyquinoline-4-oxo)-S-proline]-P1(1R,2S-VinylAcca)-COOEt, Bis HCl Salt}, N—BOC-L-tert-leucine (BOC L-tBuGly) (0.94 g,4.25 mmol), NMM (1.7 ml, 15.5 mmol) in DMF (20 mL) was added HATU (1.55g, 3.40 mmol) at rt. The reaction mixture was stirred overnite, dilutedwith 75% EtOAc-THF (300 mL), washed with pH 4.0 buffer (2×50 mL),saturated aqueous NaHCO₃ (50 mL), brine (50 mL), dried (MgSO₄), purifiedby a Biotage 40 M column (eluted with 15% to 100% EtOAc in Hexanes) tosupply the product 0.702 g (42%){BOCNH-P3(L-t-BuGly)-P2[(4R)-(7-methoxyquinoline-4-oxo)-S-proline]-P1-CO₂Etas a foam. ¹H NMR (Methanol-d₄) δ 1.06 (s, 9H), 1.22-1.32 (m, 3H), 1.28(s, 9H), 1.42-1.46 (m, 1H), 1.73 (dd, J=8, 5 Hz, 1H), 2.19-2.25 (m, 1H),2.67-2.72 (m, 1H), 3.95 (s, 3H), 4.03-4.07 (m, 1H), 4.10-4.18 (m, 2H),4.20-4.24 (m, 1H), 4.54 (d, J=12 Hz, 1H), 4.60-4.63 (m, 1H), 5.11 (dd,J=10, 2 Hz, 1H), 5.28-5.30 (m, 1H), 5.43 (m, 1H), 5.76-5.83 (m, 1H),6.50 (d, J=9 Hz, NH), 6.93 (d, J=5 Hz, 1H), 7.10 (dd, J=9, 2 Hz, 1H),7.28 (m, 1H), 7.99 (m, 1H), 8.11 (d, J=9 Hz, 1H), 8.62 (d, J=5H); LC-MSm/z 639 (retention time: 1.53 method D).

Step 8:

To a suspension of product (702 mg, 1.1 mmol) of Step 7 of Example 374{BOCNH-P3(L-t-BuGly)-P2[(4R)-7-methoxyquinoline-4-oxo)-S-proline]-P1(1R,2SVinyl Acca)-COOEt} in THF (50 mL), CH₃OH (7 mL), and H₂O (22 mL) wasadded LiOH (211 mg, 8.80 mmol). The reaction mixture was stirred for oneday, acidified to neutral pH, and concentrated in vacuo until only theaqueous layer remained. The resulting aqueous residue was acidified topH 4.0 by addition of 1.0 N aqueous HCl and then saturated with solidNaCl. This aqueous mixture was extracted repeatedly with EtOAc and THF,the combined organic solvent washed with brine (50 mL), dried (MgSO₄),filtered, and concentrated in vacuo to supply the product 631 mg (92%),BOCNH-P3(L-t-BuGly)-P2[(4R)-(7-methoxyquinoline-4-oxo)-S-proline]-P1(1R,2SVinyl Acca)-CO₂H, as a solid. ¹H NMR (Methanol-d₄) δ 1.04 (s, 9H), 1.22(s, 9H), 1.34-1.39 (m, 1H), 1.67 (dd, J=8, 5 Hz, 1H), 2.03-2.13 (m, 1H),2.43-2.49 (m, 1H), 2.67-2.73 (m, 1H), 3.96 (s, 3H), 4.00-4.05 (m, 1H),4.15-4.21 (m, 1H), 4.56-4.62 (m, 2H), 5.02 (d, J=10 Hz, 1H), 5.20 (d,J=17 Hz, 1H), 5.52 (m, 1H), 5.87-5.99 (m, 1H), 6.47 (d, J=8 Hz, 1H),6.91 (s, 1H), 7.12 (d, J=5 Hz, 1H), 7.19 (dd, J=9, 2 Hz, 1H), 7.31 (d,J=2 Hz, 1H), 8.22 (d, J=9 Hz, 1H), 8.72 (d, J=5 Hz, 1H). LC-MS(retention time: 1.44, method D), MS m/z 611 (M⁺+1).

Step 9:

To a solution of the tripeptide acid (0.120 g, 0.195 mmol) of Step 8 ofExample 374 in THF (2 mL) was added CDI (44.3 mg, 0.27 mmol) and theresulting solution was refluxed for 60 min and allowed to cool down tort. Cyclobutylsulfonamide (0.037 g, 0.273 mmol) was added in one portionbefore the addition of neat DBU (0.041 mL, 0.273 mmol). The reaction wasstirred for 24 h, another one equivalent of CDI andcyclobutylsulfonamide added and the mixture stirred 48 h more. Themixture was diluted with 50% THF/EtOAc (200 mL) and washed brinesaturated pH 4.0 buffer (30 mL), dried (MgSO₄) and concentrated invacuo. The residue was dissolved in 2 mL of 50% THF—CH₂Cl₂, 75 mg (0.39mmol) of EDAC, 48 mg (0.39 mmol) of 4-DMAP, 58 L (0.39 mmol) of DBU and53 mg (0.39 mmol) of cyclobutylsulfonamide added, and the mixturestirred 4 days. The mixture was purified by one 1000 Analtech PTLC plate(20×40 cM, eluted with 2% MeOH in CH₂Cl₂) to supply the desired productCompound 374,BOCNH-P3(L-t-BuGly)-P2[(4R)-(7-methoxyquinoline-4-oxo)-S-proline]-P1(1R,2SVinyl Acca)-CONHSO₂. Cyclobutane, as a foam 2 mg (2%): ¹H NMR(methanol-d₄) δ 1.07, 1.08 (two s (rotamers) 9H), 1.20, 1.21 (two s(rotamers) 9H), 1.41-1.48 (m, 1H), 1.64-1.70 (m, 1H), 1.72-1.91 (m, 2H),1.95-2.11 (m, 2H), 2.23-2.37 (m, 2H), 2.40-2.58 (m, 2H), 2.72-2.75 (m,1H), 4.06 (s, 3H), 4.12-4.17 (m, 2H), 4.35-4.38 (m, 1H), 4.58-4.62 (m,1H), 4.65-4.70 (m, 1H), 5.16-5.18 (m, 1H), 5.24-5.37 (m, 1H), 5.69-5.76(m, 2H), 7.40-7.46 (m, 3H), 8.35-8.40 (m, 1H), 8.92 (d, J=7 Hz, 1H).LC-MS (retention time: 1.58 method B), MS m/z 728 (M⁺+1).

Example 375 Preparation of Compound 375

Step 1:

To a solution of product (794 mg, 1.51 mmol) of Step 5 of Example 374{N—BOC-P2[(4R)-(7-methoxyquinoline-4-oxo)-S-proline]-P1(1R,2SVinylAcca)-CO₂Et} in 68 mL of 12% MeOH/THF was added a solution of 218mg (9.08 mmol) of lithium hydroxide in 30 mL of water and the mixturewas stirred 16 h. The pH was adjusted to neutral by addition of 6Naqueous HCl, concentrated until only the water remained, the solutionadjusted to pH 4 using aqueous 1N HCl and was then extracted with 50%THF-EtOAc (5×200-mL portions). The combined organic layers were dried(MgSO₄) and concentrated to provide the product 752 mg (100%){N—BOC-P2[(4R)-(7-methoxyquinoline-4-oxo)-S-proline]-P1(1R,2SVinylAcca)-CO₂H}:

¹H NMR (Methanol-d₄) 1.37-1.43 (m, 1H), 1.39 (s, 9H), 1.69-1.78 (m, 1H),2.16-2.24 (m, 1H), 2.44-2.54 (m, 1H), 2.64-2.74 (m, 1H), 3.89-3.94 (m,2H), 3.96 (s, 3H), 4.40-4.43 (m, 1H), 5.11 (d, J=10 Hz, 1H), 5.31 (d,J=17 Hz, 1H), 5.40 (m, 1H), 5.79-5.87 (m, 1H), 6.91 (s, 1H), 7.04 (d,J=6 Hz, 1H), 7.25 (dd, J=9.1, 2 Hz, 1H), 7.29 (m, 1H), 8.09 (d, J=9.1Hz, 1H), 8.66 (d, J=6 Hz, 1H). LC-MS (retention time: 1.05, method H).MS m/z 498 (M+1).

Step 2:

To a solution of product (399.5 mg, 0.668 mmol of Step 1 of Example 375{N—BOC-P2[(4R)-(7-methoxyquinoline-4-oxo)-S-proline]-P1(1R,2SVinylAcca)-CO₂H} in THF (4 mL) and CDI (434 mg, 2.68 mmol) was refluxedfor 60 min and allowed to cool down to rt. Cyclopropylsulfonamide (406mg, 3.35 mmol) was added in one portion before the addition of neat DBU(0.50 mL, 3.35 mmol). The reaction was stirred for 16 h, diluted with50% THF-EtOAc (200 mL) and washed with brine saturated pH 4.0 buffer(2×40 mL). Organic layer was dried (MgSO₄), concentrated, and purifiedover a Biotage 25M column (MeOH in CH₂Cl₂, 0% to 15%) to supply 217 mg(54%) of the desired product{N—BOC-P2[(4R)-(7-methoxyquinoline-4-oxo)-S-proline]-P1(1R,2SVinylAcca)-CONHSO₂ Cyclopropane}: ¹H NMR (Methanol-d₄) δ 1.01-1.10 (m,2H), 1.11-1.18 (m, 1H), 1.20-1.27 (m, 1H), 1.39-1.48 (m, 1H), 1.44 (s,9H), 1.87 (dd, J=8, 5 Hz, 1H), 2.01-2.38 (m, 2H), 2.57 (dd, J=14, 7 Hz,1H), 2.91-2.96 (m, 1H), 3.83-3.92 (m, 2H), 3.94 (s, 3H), 4.36-4.39 (m,1H), 5.11 (d, J=10 Hz, 1H), 5.29 (d, J=17 Hz, 1H), 5.38 (m, 1H),5.74-5.81 (m, 1H), 6.91 (d, J=5.5 Hz, 1H), 7.20 (dd, J=9.2, 2.4 Hz, 1H),7.29 (m, 1H), 8.07 (d, J=9.2 Hz, 1H), 8.60 (d, J=5.5 Hz, 1H). LC-MS(retention time: 1.28, method I). MS m/z 601 (M⁺+1)

Steps 3:

A total of product (198 mg, 0.33 mmol) of Step 2 of Example 375{BOC-P2[(4R)-(7-methoxyquinoline-4-oxo)-S-proline]-P1(1R,2S VinylAcca)-CONHSO₂ Cyclopropane} was dissolved in 4N HCl/dioxane (4 ml, 16mmol) and stirred for 2 h at rt. The reaction mixture was concentratedto supply the crude product as a tan solid which was used immediately inthe next reaction.

Step 4:

The crude product of Step 3 of Example 375{HN-P2[(4R)-(7-methoxyquinoline-4-oxo)-S-proline]-P1(1R,2S VinylAcca)-CONHSO₂ Cyclopropane, Bis HCl Salt} was suspended in 10 mL ofdichloromethane. To this mixture was added N—BOC-L-tert-leucine (BOCL-tBuGly) [120 mg, 0.52 mmol], HOAT (30 mg, 0.20 mmol), DIPEA (0.29 ml,1.65 mmol), and HATU (160 mg, 0.43 mmol) at rt. The reaction mixture wasstirred for 16 h, diluted with 50% EtOAc-THF (300 mL), washed with brinesaturated pH 4.0 buffer (3×50 mL), dried (MgSO₄), concentrated. Theresidue was purified by a Isco 35 g column (eluted with 0% to 15% MeOHin CH₂Cl₂) to supply the product (130.1 mg, 47%) as a Hunning's basesalt (Compound 375): ¹H NMR (methanol-d₄) δ ppm 1.00-1.48 (m, 29H), 1.47(s, 9H), 1.89 (m, 1H), 2.26 (m, 1H), 2.36 (m, 1H), 2.69 (m, 1H), 2.97(m, 1H), 3.25 (q, J=7.43 Hz, 2H), 3.74 (m, 2H), 3.97 (s, 3H), 4.10 (m,1H), 4.23 (dd, J=19.68, 9.92 Hz, 1H), 4.57 (m, 2H), 5.15 (m, 1H), 5.31(m, 1H), 5.50 (s, 1H), 5.77 (m, 1H), 7.01 (t, J=5.34 Hz, 1H), 7.16 (d,J=9.16 Hz, 1H), 7.31 (d, J=1.83 Hz, 1H), 8.14 (m, 1H), 8.67 (d, J=5.49Hz, 1H).

LC-MS (retention time: 1.49 Method d), MS m/z 714 (M⁺+1).

Example 376 Preparation of Compound 376

Step 1:

Of Compound 375 (130 mg) was dissolved in EtOAc, washed one more timewith pH 4 buffer, brine and then dried (MgSO₄). The crude mixture waspurified over two 1000 PTLC plate from Analtech (20×40 cm, eluted with3% MeOH in CH₂Cl₂) to afford the product (Compound 376) 54 mg (23% yieldfrom the tripeptide acid): ¹H NMR (Methanol-d₄) δ 0.88-1.00 (m, 2H),1.01-1.14 (m, 2H), 1.03 (s, 9H), 1.25 (s, 9H), 1.34 (dd, J=9, 5 Hz, 1H),1.81-1.89 (m, 1H), 2.06-2.13 (m, 1H), 2.45-2.50 (m, 1H), 2.65-2.75 (m,1H), 3.91 (s, 3H), 3.98-4.11 (m, 1H), 4.21-4.22 (m, 1H), 4.46-4.50 (m,1H), 4.54-4.57 (m, 1H), 4.97-5.02 (m, 1H), 5.14-5.22 (m, 1H), 5.33-5.41(m, 1H), 5.81-5.99 (m, 1H), 6.87-6.95 (m, 1H), 7.06-7.09 (m, 1H), 7.25(m, 1H), 8.07-8.10 (m, 1H), 8.59 (d, J=5.2 Hz, 1H). HRMS m/z (M−H)⁻calcd. for C₃₅H₄₆N₅O₉S: 712.3016, found: 712.3024; LC-MS m/e 714(retention time: 1.42, method I).

Example 377 Preparation of Compound 377

Step 1:

A total of 1.0 mmol the product of Step 2 of Example 375 {The Bis HClSalt of HN-P2[(4R)-(7-methoxyquinoline-4-oxo)-S-proline]-P1(1R,2S VinylAcca)-CONHSO₂ Cyclopropane}suspended in 20 mL of dichloromethane wasadded 352 mg (1.30 mmol) of 2-(S)-tert-butoxycarbonylamino-8-nonenoicacid purchased from RSP Amino Acids, HOAT (82 mg, 0.60 mmol), DIPEA(0.74 ml, 5.0 mmol), and HATU (494 mg, 1.30 mmol) at rt. The reactionmixture was stirred 16 h, and the majority of the CH₂Cl₂ removed invacuo. The mixture was diluted with saturated pH 4.0 buffer (150 mL),and extracted into EtOAc (4×200 mL). The combined organic layers weredried (MgSO₄), concentrated. The residue was purified over a Biotage 40Mcolumn (eluted with 0% to 15% MeOH in CH₂Cl₂) to afford the product(Compound 377) 574 mg (76%): LC-MS m/z 754 (retention time: 1.64, methodI).

Example 378 Preparation of Compound 378

Step 1:

A total of 0.34 mmol of the product of Step 2 of Example 375 {The BisHCl Salt of HN-P2[(4R)-(7-methoxyquinoline-4-oxo)-S-proline]-P1(1R,2SVinyl Acca)-CONHSO₂ Cyclopropane} was suspended in 3 mL ofdichloromethane. To this mixture was added N—BOC-L-Valine (L-Val) (120mg, 0.55 mmol), HOAT (30 mg, 0.20 mmol), DIPEA (0.29 ml, 1.65 mmol), andHATU (160 mg, 0.43 mmol) at rt. The reaction mixture was stirred 16 h,diluted with saturated pH 4.0 buffer (150 mL), and extracted into EtOAc(3×200 mL). The combined organic layers were washed with brine, dried(MgSO₄), and concentrated. The residue was purified over an Isco 35 gcolumn (MeOH in CH₂Cl₂: 0% to 15%). This material was further purifiedover two δ PTLC plate from Analtech (20×40 cm, eluted with 3% MeOH inCH₂Cl₂) to afford the product 104.1 mg (44%), Compound 378: HRMS m/z(M−H)⁻ calcd. for C₃₄H₄₄N₅O₉S: 698.2860, found: 698.2865. LC-MS m/e 700(retention time: 1.60, method D).

Example 379 Preparation of Compound 379

Step 1:

To a suspension of 2-picolic acid (3.73 g, 30.3 mmol) and2-amino-4-methoxybenzophenone (5.0 g, 30.3 mmol) at −30° C. dissolved inpyridine (150 mL) was added POCl₃ (3.7 mL, 45.4 mmol) in 5 min. thereaction mixture was stirred for 3 hr at the temperature, and stirred atrt overnite. The reaction mixture was poured into cold water andextracted with EtOAc (3×). The combined extract was dried to provide theproduct (7.67 g, 93%): ¹H NMR (methanol-d₄) δ ppm 2.65 (s, 3H), 3.92 (s,3H), 6.78 (m, 1H), 7.60 (m, 1H), 8.00 (m, 1H), 8.06 (m, 1H), 8.21 (d,J=7.63 Hz, 1H), 8.59 (t, J=2.29 Hz, 1H), 8.76 (d, J=3.97 Hz, 1H). LC-MS(retention time: 1.56, Method D), MS m/z 271 (M⁺+1).

Step 2:

To a suspension of Pyridine-2-carboxylic acid(2-acetyl-5-methoxy-phenyl)-amide (2.90 g, 10.7 mmol) in THF (50 mL) wasadded t-BuOK/THF (1M, 24 mL, 24 mmol). The reaction mixture was heatedat 70° C. for 3 h and stirred overnite. The solvent was removed the invacuo. Cold water was added to the residue and adjusted pH to 4.6 withaqueous 1.0 N HCl, filtered. The solid residue was purified over aBiotage 65M column (MeOH/CH₂Cl₂: 0-15%) to provide the product (2.26 g,84%): LC-MS (retention time: 1.19, Method D), MS m/z 253 (M⁺+1).

Step 3:

A mixture of 7-Methoxy-2-pyridin-2-yl-quinolin-4-ol (2.2 g, 8.71 mmol)in POCl₃ (92 mL) was refluxed for 3 h and then removed the solvent invacuo. Ice water was added to the residue, adjusted the pH>10 with 1.0 NNaOH, and extracted with EtOAc (2×). The combined extract was washedwith water, brine, dried (MgSO₄), removed solvent to supply the productas a yellow solid (89%, 2.1 g): DMSO-D6) δ ppm 3.97 (s, 3H), 7.40 (dd,J=9.16, 2.44 Hz, 1H), 7.53 (m, 1H), 8.01 (m, 1H), 8.09 (d, J=9.16 Hz,1H), 8.46 (s, 1H), 8.56 (d, J=7.93 Hz, 1H), 8.74 (d, J=3.97 Hz, 1H).LC-MS (retention time: 1.50, Method D), MS m/z 271 (M⁺+1).

Step 4:

To a solution of N-Boc-4-hydroxyproline (1.6 g, 6.7 mmol) in DMSO (20mL) was added t-BuOK (1.9 g, 16.8 mmol). The generated mixture wasstirred for 1.5 h and 4-Chloro-7-methoxy-2-pyridin-2-yl-quinoline (2.0g, 7.4 mmol) and DMSO (10 mL) were added. The reaction mixture wasstirred for 38 h, diluted with cold water and extracted with EtOAc/ether(1/4, 2×). the aqueous layer was acidified to pH 4 and extracted withEtOAc/THF (5×). the combined extract was dried (Na₂SO₄/MgSO₄), removedthe solvent in vacuo and the residue was purified by preparative HPLC(0-80% solvent B) to provide the product (1.6 g, 50%): LC-MS (retentiontime: 1.23, Method I), MS m/z 466 (M+11).

Step 5:

A solution of product (0.21 g, 0.65 mmol) of Step 4 of Example 379{N-boc-(1R,2S)-1-amino-2-vinylcyclopropane carboxylic acid ethyl ester}in HCl/dioxane (4M, 5 mL, 20 mmol) was stirred for 3 h, and the solventwas removed in vacuo. To the residue was added CH₂Cl₂ (10 mL),diisopropylethylamine (0.4 mL, 3.23 mmol), HOBT (0.20 g, 1.35 mmol),Boc-(4R)-(2-cyclopropyl-7-methoxy-quinoline-4-oxo)-S-proline (0.20 g,0.5 mmol) and HATU (0.415 g, 1.07 mmol). The reaction mixture wasstirred overnite and diluted with pH 4.0 buffer, extracted with EtOAc.The extract was dried (MgSO₄) and purified by Biotage 40 M column usingMeOH/CH₂Cl₂ (0 to 15%) as eluent to provide the product (204.7 mg, 70%):¹H NMR (methanol-d₄) δ ppm 0.64 (m, 1H), 0.96 (m, 2H), 1.33 (m, 8H),1.39 (m, 9H), 1.90 (m, 2H), 2.18 (m, 1H), 2.54 (m, 1H), 2.81 (m, 1H),4.01 (m, 5H), 4.44 (d, J=28.99 Hz, 1H), 5.08 (m, 1H), 5.31 (m, 1H), 5.57(s, 1H), 6.03 (m, 1H), 6.94 (s, 1H), 7.27 (d, J=8.24 Hz, 1H), 7.64 (m,1H), 7.92 (m, 1H), 8.14 (m, 2H), 8.66 (s, 1H), 8.74 (s, 1H).

Step 6:

A slurry of P2Boc-(4R)-(7-methoxy-2-Pyridin-2-yl-quinoline-4-oxo)-S-proline]-P1(1R,2SVinyl Acca)-CONHSO₂(1-cyclopropylmethylcyclopropan-1-yl) (Step 5,Example 379) (203 mg, 0.3 mmol) in 4M HCl/dioxane (3.5 mL, 14 mmol) wasstirred for 2 h, removed the solvent in vacuo. To the residue was addedCH₂Cl₂ (2 mL), diisopropylethylamine (0.63 mL, 3.6 mmol),Boc-L-tert-leucine (83 mg, 0.36 mmol), HOAt (41 mg, 0.3 mmol), and HATU(148 mg, 0.39 mmol). The reaction mixture was stirred at rt for 7 h andremoved the solvent in vacuo. The residue was purified by preparativeHPLC (35-85% solvent B) to provide the desired product (Compound 379)25.1 mg (11%): ¹H NMR (methanol-d₄) δ ppm −0.05 (m, 1H), 0.30 (m, 1H),0.66 (m, 1H), 0.91 (m, 2H), 1.05 (s, 9H), 1.28 (s, 9H), 1.67 (m, 8H),2.15 (m, 1H), 2.58 (m, 1H), 2.77 (m, 1H), 3.96 (s, 3H), 4.19 (d, J=40.25Hz, 2H), 4.51 (d, J=16.47 Hz, 2H), 4.95 (m, 1H), 5.15 (m, 1H), 5.53 (s,1H), 5.89 (dd, J=16.65, 9.33 Hz, 1H), 7.09 (d, J=8.42 Hz, 1H), 7.43 (d,J=1.83 Hz, 1H), 7.50 (m, 1H), 7.82 (s, 1H), 7.99 (m, 1H), 8.10 (d,J=9.15 Hz, 1H), 8.48 (d, J=7.68 Hz, 1H), 8.72 (s, 1H). LC-MS (retentiontime: 1.59, Method I), MS m/z 791 (M⁺+1).

Example 380 Preparation of Compound 380

Step 1:

The starting material in Scheme 1 of the present example was prepared bycoupling of the product of step 3 in Example 11 of section B with theamino terminus of P1(1R,2S Vinyl Acca)-COOEt. A slurry of said couplingproduct, P2 Boc-(4R)-(6-methoxy-isoquinoline-1-oxo)-S-proline]-P1(1R,2SVinyl Acca)-COOEt (7.88 g, 14.99 mmol) in 4M HCl/dioxane (120 mL, 480mmol) was stirred for 2 h, removed the solvent in vacuo and azeotropedwith dry dioxane. To the residue was added DMF (75 mL),N-methylmorpholine (6.27 mL, 57.07 mmol), Boc-L-tert-leucine (5.20 g,22.49 mmol), and HATU (8.53 g, 22.49 mmol). The reaction mixture wasstirred at rt overnite and worked up by pouring the reaction mixtureinto ice water and adjusted to pH 5 with aqueous 1.0 N HCl and extractedwith EtOAc. The extract was washed with NaHCO₃ (aq.), brine, dried(MgSO₄) and concentrated. The residue was purified over Biotage 65Mcolumn (EtOAc-hexanes: 5-100%) to provide the product (8.07 g, 84%):Retention time: 1.88 method I) MS m/z 639 (M⁺+1).

Step 2:

To a suspension of the product (4.0 g, 6.26 mmol) of Step 1 of Example384{Boc-NH-P3(L-tert-BuGly)-P2[(4R)-(6-methoxyl-isoquinoline-1-oxo)-S-proline]-P1(1R,2SVinyl Acca)-COOEt} in THF (250 mL), CH₃OH (31 mL), and H₂O (125 mL) wasadded LiOH (2.4 g, 100.2 mmol). The reaction mixture was stirred forovernite and then adjusted to pH 7 with aqueous 1.0 N HCl. The organicsolvents were removed in vacuo. The aqueous residue was acidified to pH4 and extracted with EtOAc (2×).

The combined organic solvent was dried (Na₂SO₄/MgSO₄), and concentratedin vacuo to supply the product (3.79 g, 99%): ¹H NMR (methanol-d₄) ppm1.05 (s, 9H), 1.25 (m, 1H), 1.29 (s, 9H), 1.46 (m, 1H), 1.72 (dd,J=8.24, 5.19 Hz, 1H), 2.23 (q, J=8.55 Hz, 1H), 2.68 (dd, J=13.89, 7.78Hz, 1H), 3.94 (s, 3H), 4.05 (dd, J=11.60, 3.05 Hz, 1H), 4.23 (d, J=8.85Hz, 1H), 4.46 (d, J=11.60 Hz, 1H), 4.63 (t, J=8.39 Hz, 1H), 5.10 (d,J=10.38 Hz, 1H), 5.29 (d, J=17.40 Hz, 1H), 5.85 (m, 2H), 7.10 (d, J=9.16Hz, 1H), 7.19 (s, 1H), 7.26 (d, J=5.49 Hz, 1H), 7.91 (d, J=5.80 Hz, 1H),8.12 (d, J=9.16 Hz, 1H). Retention time: 1.81 method I) MS m/z 611(M⁺+1).

Step 3:

A solution of CDI (0.052 g, 0.32 mmol) and the product (0.130 g, 0.21mmol) of Step 2 of Example 384{BOCNH-P3(L-t-BuGly)-P2[(4R)-6-methoxy-sioquinoline-1-oxo)-S-proline]-P1(1R,2SVinyl Acca)-CO₂H} in THF (2 mL) was refluxed for 60 min and allowed tocool down to rt. Cyclobutanesulfonamide (0.043 g, 0.32 mmol) was addedfollowed by the addition of a solution of neat DBU (0.048 mL, 0.32mmol). The reaction was stirred for overnite, then filtered throughsyringe filter and purified by preparative HPLC (30% to 100% solvent B)to provide the desired product 0.1422 mg (92%): ¹H NMR (methanol-d₄) δppm 1.04 (s, 9H), 1.26 (d, J=13.43 Hz, 9H), 1.39 (m, 1H), 1.85 (dd,J=7.63, 5.19 Hz, 1H), 1.98 (m, 2H), 2.26 (m, 4H), 2.50 (m, 2H), 2.61 (m,1H), 3.92 (s, 3H), 4.05 (m, 1H), 4.24 (m, 1H), 4.33 (m, 1H), 4.43 (d,J=11.60 Hz, 1H), 4.52 (m, 1H), 5.13 (m, 1H), 5.30 (m, 1H), 5.71 (m, 1H),5.82 (s, 1H), 7.08 (d, J=8.85 Hz, 1H), 7.18 (s, 1H), 7.24 (d, J=5.80 Hz,1H), 7.88 (m, 1H), 8.08 (d, J=9.16 Hz, 1H). Retention time: 1.89 methodI) MS m/z 728 (M⁺+1).

Step 4:

Example 380, Step 3 (0.196 mg, 0.27 mmol){(BOCNH-P3(L-t-BuGly)-P2-(4R)-(6-methoxy-isoquinoline-1-oxo)-S-proline]-P1(1R,2SVinyl Acca)-CONHSO₂ Cyclobutyl} was dissolved in HCl/dioxane (5 mL; 20mmol) and was stirred for 2 h at rt. Removed the solvent in vacuo tosupply the titled product 100% (0.1887 g) which was ready to next step.

Step 5:

To a mixture of the product (0.037 g, 0.053 mmol) of Step 4 of Example380 {HCl salt ofNH₂—P3(L-t-BuGly)-P2-(4R)-(6-methoxy-isoquinoline-1-oxo)-S-proline]-P1(1R,2SVinyl Acca)-CONHSO₂ Cyclobutyl} and diisopropylethylamine (0.046 mL),0.26 mmol) in CH₂Cl₂ (2 mL) was added cyclopentyl chloroformate (0.7 M,0.151 mL, 0.069 mmol). The reaction mixture was stirred overnite andpurified by preparative HPLC (30% to 100% solvent B) to provide thedesired product (Compound 380) (0.0303 g, 77%): ¹H NMR (methanol-d₄) δppm 1.03 (s, 9H), 1.48 (m, 9H), 1.86 (dd, J=8.24, 5.49 Hz, 1H), 1.99 (m,2H), 2.27 (m, 4H), 2.51 (m, 2H), 2.60 (dd, J=13.89, 6.87 Hz, 1H), 3.92(s, 3H), 4.05 (dd, J=12.21, 3.97 Hz, 1H), 4.32 (m, 2H), 4.41 (d, J=11.90Hz, 1H), 4.53 (m, 1H), 4.69 (m, 1H), 5.12 (d, J=10.38 Hz, 1H), 5.29 (d,J=17.09 Hz, 1H), 5.71 (m, 1H), 5.83 (s, 1H), 7.11 (d, J=9.46 Hz, 1H),7.19 (s, 1H), 7.25 (d, J=5.80 Hz, 1H), 7.88 (d, J=5.80 Hz, 1H), 8.08 (d,J=9.16 Hz, 1H). retention time: 1.85 method H), MS m/z 740 (M⁺+1)

Example 381 Preparation of Compound 381

Step 1:

To a mixture of the product (0.037 g, 0.053 mmol) of Step 4 of Example380 {HCl salt ofNH₂—P3(L-t-BuGly)-P2-(4R)-(6-methoxy-isoquinoline-1-oxo)-S-proline]-P1(1R,2SVinyl Acca)-CONHSO₂ Cyclobutyl} and isopropylethylamine (0.046 mL), 0.26mmol) in CH₂Cl₂ (2 mL) was added new-pentyl chloroformate (0.012 mL,0.069 mmol). The reaction mixture was stirred overnite and directlypurified by preparative HPLC (30% to 100% solvent B) to provide thedesired product (Compound 381) (0.0252 g, 64%): ¹H NMR (methanol-d₄) δppm 0.84 (s, 9H), 1.05 (s, 9H), 1.40 (m, 1H), 1.86 (m, 1H), 2.00 (m,2H), 2.28 (m, 4H), 2.51 (m, 2H), 2.57 (m, 1H), 3.39 (d, J=10.07 Hz, 1H),3.55 (d, J=10.38 Hz, 1H), 3.92 (s, 3H), 4.05 (m, 1H), 4.33 (m, 2H), 4.41(d, J=11.29 Hz, 1H), 4.53 (m, 1H), 5.12 (d, J=10.07 Hz, 1H), 5.29 (d,J=17.09 Hz, 1H), 5.71 (m, 1H), 5.82 (s, 1H), 7.10 (d, J=9.16 Hz, 1H),7.19 (s, 1H), 7.25 (d, J=5.80 Hz, 1H), 7.88 (d, J=5.80 Hz, 1H), 7.97 (s,1H), 8.07 (d, J=8.85 Hz, 1H). retention time: 1.89 method H), MS m/z 742(M⁺+1).

Example 382 Preparation of Compound 382

Step 1:

To a mixture of the product (0.037 g, 0.053 mmol) of Step 4 of Example380 {HCl salt ofNH₂—P3(L-t-BuGly)-P2-(4R)-(6-methoxy-isoquinoline-1-oxo)-S-proline]-P1(1R,2SVinyl Acca)-CONHSO₂ Cyclobutyl} and diisopropylethylamine (0.046 mL),0.26 mmol) in CH₂Cl₂ (2 mL) was added di-t-amyl dicarbonate (0.0169 g,0.069 mmol). The reaction mixture was stirred overnite and directlypurified by HPLC (30% to 100% solvent B) to provide the desired product(Compound 382) (0.0175 g, 44%): ¹H NMR (methanol-d₄) δ ppm 0.79 (t,J=6.87 Hz, 3H), 1.04 (s, 8H), 1.21 (s, 3H), 1.23 (s, 3H), 1.41 (m, 2H),1.64 (m, 2H), 1.83 (m, 1H), 2.00 (m, 2H), 2.26 (m, 4H), 2.51 (m, 2H),2.60 (m, 1H), 3.92 (s, 3H), 4.07 (m, 1H), 4.24 (m, 1H), 4.33 (m, 1H),4.43 (d, J=11.60 Hz, 1H), 4.52 (m, 1H), 5.13 (m, 1H), 5.29 (m, 1H), 5.71(m, 1H), 5.82 (s, 1H), 7.09 (d, J=8.85 Hz, 1H), 7.18 (s, 1H), 7.25 (d,J=5.49 Hz, 1H), 7.88 (d, J=5.80 Hz, 1H), 8.08 (d, J=8.85 Hz, 1H).retention time: 1.90, method H), MS m/z 742 (M⁺+1).

Example 383 Preparation of Compound 383

Step 1

To a mixture of the product (0.037 g, 0.053 mmol) of Step 4 of Example380 {HCl salt ofNH₂—P3(L-t-BuGly)-P2-(4R)-(6-methoxy-isoquinoline-1-oxo)-S-proline]-P1(1R,2SVinyl Acca)-CONHSO₂ Cyclobutyl} and isopropylethylamine (0.046 mL), 0.26mmol) in CH₂Cl₂ (2 mL) was added t-butyl-isocyanate (0.008 mL, 0.069mmol). The reaction mixture was stirred overnite and directly purifiedby preparative HPLC (30% to 100% solvent B) to provide the desiredproduct (Compound 383) (0.024 g, 62%): ¹H NMR (methanol-d₄) δ ppm 1.05(s, 9H), 1.19 (s, 9H), 1.37 (m, 1H), 1.85 (dd, J=8.09, 5.34 Hz, 1H),2.00 (m, 2H), 2.26 (m, 4H), 2.50 (m, 2H), 2.58 (m, 1H), 3.92 (s, 3H),4.06 (m, 1H), 4.32 (m, 2H), 4.49 (m, 2H), 5.11 (d, J=10.38 Hz, 1H), 5.27(d, J=17.40 Hz, 1H), 5.69 (m, 1H), 5.83 (s, 1H), 7.08 (dd, J=9.16, 2.44Hz, 1H), 7.17 (d, J=2.44 Hz, 1H), 7.24 (d, J=5.80 Hz, 1H), 7.87 (d,J=6.10 Hz, 1H), 8.12 (d, J=8.85 Hz, 1H). retention time: 1.77, methodH), MS m/z 727 (M⁺+1).

Example 384 Preparation of Compound 384

Step 1:

A suspension of diisopropylethylamine (0.031 mL, 0.018 mmol),N,N′-disuccinimidyl carbonate (0.0274 g, 0.107 mmol) and the product(0.050 g, 0.0714 mmol) of Step 4 of Example 380 {HCl salt ofNH₂—P3(L-t-BuGly)-P2-(4R)-(6-methoxy-isoquinoline-1-oxo)-S-proline]-P1(1R,2SVinyl Acca)-CONHSO₂ Cyclobutyl} in THF (2 mL) was sonicated at 80° C.for 15 min. KH (0.046 g, 1.14 mmol) and 1-methylcyclopentanol (0.079 mL,0.714 mmol) was added. The reaction mixture was stirred for 20 min andworked up by diluting with cold water, adjusted pH to 4, extracted withEtOAc. The extract was dried (MgSO₄) and the residue was purified bypreparative HPLC (30% to 100% solvent B) to provide the desired product(Compound 384) (0.018 g, 33%): (methanol-d₄) δ ppm 1.04 (s, 9H),1.29-1.79 (m, 10H), 1.84 (m, 2H), 1.99 (m, 3H), 2.26 (m, 4H), 2.49 (m,2H), 2.60 (dd, J=13.73, 7.02 Hz, 1H), 3.92 (s, 3H), 4.05 (dd, J=11.29,2.44 Hz, 1H), 4.26 (s, 1H), 4.32 (m, 1H), 4.44 (d, J=11.90 Hz, 1H), 4.52(m, 1H), 5.12 (d, J=10.07 Hz, 1H), 5.28 (d, J=16.79 Hz, 1H), 5.71 (m,1H), 5.82 (s, 1H), 7.10 (d, J=8.85 Hz, 1H), 7.19 (s, 1H), 7.26 (d,J=5.80 Hz, 1H), 7.88 (d, J=5.80 Hz, 1H), 8.08 (d, J=9.16 Hz, 1H). LC-MSretention time: 1.91 method H), MS m/z 754 (M+1).

Example 385 Preparation of Compound 385

Step 1:

A suspension of 2-cyanomethyl-4-methoxy-benzoic acid methyl ester (1.9 gand TsOH. H₂O (0.15 g, mmol) in morpholine 5 mL) was refluxed for 4 hand removed the solvent in vacuo. The residue was recrystalyzed fromEtOAc/hexanes with drops of MeOH to provide the product (0.43 g, 17%):LC-MS retention time: 1.07 method H), MS m/z 266 (M⁺+1).

Step 2:

A mixture of 6-methoxy-3-morpholin-4-yl-isoquinolin-1-ol (0.298 g, 1.15mmol) in POCl₃ (20 mL) was refluxed for 2 h, removed the solvent invacuo and cold water was added. The pH was adjusted to >11 by additionof 1.0 N NaOH. The aqueous layer was extracted with EtOAc. The extractwas dried (MgSO₄), removed the solvent in vacuo to provide the predate(0.299 g, 94%): LC-MS retention time: 1.68 method H), MS m/z 279 (M⁺+1).

Step 3:

A mixture of 1-Chloro-6-methoxy-3-morpholin-4-yl-isoquinoline (0.050 g,0.18 mmol) and tetrabutyl phosphorium hydrogen difluoride (0.8 g, 2.8mmol) [Synlett 1992, (4), 345-6] was heated at 140° C. in microwave for10 min. the reaction mixture was diluted with EtOAc and filtered throughan ISCO 25 g precolumn with a layer of silicon gel on the top, removedthe solvent to provide the product (0.037 mg, 77%): ¹H NMR(CHLOROFORM-D) δ ppm 3.48 (m, 4H), 3.84 (m, 4H), 3.89 (s, 3H), 6.46 (d,J=1.22 Hz, 1H), 6.85 (s, 1H), 6.90 (dd, J=9.16, 2.44 Hz, 1H), 7.82 (d,J=8.85 Hz, 1H). LC-MS retention time: 1.56 method H), MS m/z 263 (M⁺+1).

Step 4:

A mixture of 1-fluoro-6-methoxy-3-morpholin-4-yl-isoquinoline (0.037 g,0.14 mmol), LaCl₃ (0.020 g, 0.8 mmol), t-BuOK (1M/THF, 0.32 mL, 0.32mmol), andBoc-NH—P3(L-tert-BuGly)-P2[(4R)-4-hydroxyl-5-proline]-P1(1R,2S VinylAcca)-CONHSO₂ Cyclopropane (0.045 g, 0.08 mmol) in THF (3 mL) wasstirred for 3 days. The reaction mixture was diluted with methanolfiltered through syringe filter and purified by preparative HPLC toprovide the product as a pale yellow foam (0.0158 g, 24%): ¹H NMR(methanol-d₄) δ ppm 1.03 (s, 9H), 1.24 (m, 4H), 1.31 (s, 9H), 1.43 (m,2H), 1.88 (m, 1H), 2.24 (m, 2H), 2.59 (dd, J=13.43, 6.71 Hz, 1H), 2.94(m, 1H), 3.47 (m, 4H), 3.83 (m, 4H), 3.86 (s, 3H), 4.08 (m, 1H), 4.28(s, 1H), 4.48 (m, 1H), 5.12 (d, J=10.38 Hz, 1H), 5.29 (d, J=16.48 Hz,1H), 5.76 (m, 2H), 6.74 (d, J=9.16 Hz, 1H), 6.94 (s, 1H), 7.85 (d,J=8.85 Hz, 1H), 9.19 (s, 1H). retention time: 1.86 method H), MS m/z 799(M⁺+1).

Example 386 Preparation of Compound 386

Compounds 386 was prepared using the methods described herein.

Section I

All compounds in section I were analyzed by the LC/MS methodology, whichhas the following conditions.

Method A: Xterra C18 S7 3.0×50 mm

Gradient: 100% solvent A/0% solvent B to 0% solvent A/100% solvent B

Gradient time: 3 min.

Hold time: 1 min.

Flow rate: 4 mL/min.

Detector Wavelength: 220 nm

Solvent A: 10% MeOH/90% H₂O/0.1% TFA

Solvent B: 10% H₂O/90% MeOH/0.1% TFA

Example 410 Preparation of Compound 410

Step 1:

To a solution of Boc-L-hydroxyproline (0.723 g, 3.13 mmol) in DMSO,KO^(t)Bu (0.808 g, 7.2 mmol) was added under a nitrogen atmosphere. Thesuspension was stirred at room temperature for 1.5 hours, and4-chloro-6-methyl-2-(trifluoromethyl)quinoline (0.916 g, 3.75 mmol) wasadded in two portions. The mixture was stirred at room temperature forthree hours, and 1.3 equivalents of HCl (1N) was used to neutralize thereaction. Buffer solution of pH 4.0 was added and the pH was adjusted topH 4-5. The aqueous layer was extracted with ethyl acetate, (3×25 mL)and the combined organic layers were washed with brine (20 mL) and driedover MgSO₄ to yield the titled compound as a white solid (crude yieldnot calculated). The crude product was taken into the next step.

LC/MS rt-min (MH⁺): 2.48 (441.5) (method A).

Step 2:

A solution of the crude product from Step1,4-(6-methyl-2-trifluoromethyl-quinolin-4-yloxy)-pyrrolidine-1,2-dicarboxylicacid 1-tert-butyl ester (assumed 3.13 mmol), in THF (10 mL) and methanol(10 mL) was cooled to 0° C. TMSCN₂ 2M in hexanes (˜1.3 eq) was slowlyadded to the stirring solution under a nitrogen atmosphere until gas wasno longer emitted from the solution. The fully reacted solution was thenconcentrated in vacuo, and purification by a Biotage 40M column (eluted10%-30% ethyl acetate in hexanes) afforded the pure titled compound as awhite foam (976 mg, 69% over Step 1&2)

LC/MS rt-min (MH⁺): 2.60 (477) (method A).

Step 3:

A solution of the product from Step 2 (0.976 g, 2.15 mmol) in DCM (7 mL)and TFA (6.62 mL) was stirred at room temperature for one hour. Thesolvent was removed in vacuo and the residue was suspended in 1N HCl indiethyl ether (8 mL), gently stirred, and concentrated in vacuo. Thisprocedure was repeated and the resulting product was placed on an oilpump overnight to yield a white solid in quantitative yield.

¹H NMR: (DMSO-d₆) δ 2.50 (s, 3H), 2.57-2.6 (m, 1H), 2.66-2.71 (m, 1H),3.62-3.65 (br d, J=15 Hz, 1H), 3.80-3.81 (m, 4H), 4.8 (br s, 1H), 5.7(s, 1H), 7.46 (s, 1H), 7.72-7.75 (d, J=7.5 Hz, 1H), 7.98-7.8 (d, J=8.5Hz, 1H), 8.24 (s, 1H), 9.54 (br s, 1H); LC/MS rt-min (MH⁺): 1.61 (355)(method A).

Step 4:

The product from Step 3 (assumed quantitative yield, 2.746 mmol) wasadded to a solution of BOC-t-Butyl-L-glycine (0.635 g, 2.746 mmol) inDCM (20 mL) under a nitrogen atmosphere. This step was followed by theaddition of HOBt (0.408 g, 3.02 mmol), DIPEA (3.35 mL, 19.2 mmol), andHBTU (1.56 g, 4.12 mmol). A peach colored solution immediately resultedand the reaction was left to stir at room temperature overnight. 10 mLDCM was added to the completed reaction in order to increase the volume,and the reaction was quenched with pH 4.00 buffer solution (25 mL). Themixture was acidified to a pH of 4.5 using 1N HCl, and the aqueous phasewas extracted with DCM (3×20 mL). The organic phase was washed twicewith pH 4.00 buffer solution (20 mL), saturated NaOH (25 mL), and brine(20 mL), and then dried with MgSO₄. The resulting solution wasconcentrated in vacuo and purified by a Biotage 40M column (eluted10%-40% ethyl acetate in hexanes). This purification afforded the puretitled compound as a white solid (1.11 g, 89%).

¹H NMR: (DMSO-d₆) δ 0.96-1.02 (rotamers, 3:2, s, 18H), 2.27-2.33 (m,1H), 2.50 (s, 3H), 2.68-2.72 (m, 1H), 3.67 (s, 3H), 4.02-4.04 (m, 1H),4.43-4.45 (br d, J=15 Hz, 2H), 4.58-4.61 (t, 1H), 5.60 (br s, 1H),6.72-6.74 (br d, J=15 Hz, 1H), 7.38 (s, 1H), 7.68-7.73 (m, 1H),7.95-7.97 (m, 2H);

LC/MS rt-min (MH⁺): 2.61 (590) (method A).

Step 5:

LiOH (0.138 g, 5.78 mmol) was dissolved in water (10 mL) by heating andsonication. The LiOH solution and MeOH (10 mL) were added to a solutionof the pure material from Step 4 (1.09 g, 1.93 mmol) in THF (10 mL). Themixture immediately turned a vivid blue color. The reaction was left tostir at room temperature for 3 hours and was then acidified with 1N HCl(5.78 mL, 5.78 mmol). The reaction was quenched with pH 4.00 buffersolution and the pH was adjusted to pH 4.5 using 1N aqueous NaOH. Theaqueous layer was extracted with EtOAc (3×25 mL), washed with brine (20mL), and dried over MgSO₄. The filtered solution was concentrated invacuo and left on a vacuum line overnight. The crude product (957 mg,90% yield) was taken into the next step.

LC/MS rt-min (MH⁺): 2.51 (577) (method A).

Step 6:

The crude product from Step 5 (60 mg, 0.11 mmol) was dissolved in DCM (5mL) and cyclopropanesulfonic acid (1(R)-amino-2(S)-vinyl-cyclopropanecarbonyl)-amide hydrochloride salt (Example 1,Step 8) (0.029 g, 0.11 mmol) was added. DIPEA (0.094 mL, 0.541 mmol),and then HATU (0.0575, 0.151 mmol) were added under a nitrogenatmosphere. The reaction was left to stir at room temperature for about8 hours. 10 mL DCM was added to the solution in order to increase thevolume, and the reaction was quenched with pH 4.00 buffer solution (10mL). The mixture was acidified using 1N HCl to a pH of 4-5, and theaqueous phase was extracted with DCM (3×20 mL). The organic phase waswashed twice with pH 4.00 buffer solution (10 mL) and brine (10 mL), andthen dried over MgSO₄. The resulting solution was concentrated in vacuoand purified by a Biotage 12M column (eluted 10%-40% acetone inhexanes). This purification afforded Compound 410 as a white powder (29mg, 35%).

¹H NMR: (DMSO-d₆) δ 0.976-1.12 (m, 24H), 1.36-1.39 (m, 1H), 1.70-1.72(m, 1H), 2.15-2.25 (m, 2H), 2.50-2.52 (m, 4H), 2.91-2.96 (m, 1H),3.97-4.01 (m, 2H), 4.40-4.47 (m, 2H), 5.09-5.11 (d, J=10 Hz, 1H),5.21-5.24 (d, J=15 Hz, 1H), 5.59-5.66 (m, 2H), 6.65-6.67 (d, NH), 7.43(s, 1H), 7.72-7.74 (d, J=10 Hz, 1H), 7.90 (s, 1H), 7.98-8.0 (d, J=10 Hz,1H), 8.87 (s, NH), 10.35 (s, NH);

LC/MS rt-min (MH⁺): 2.65 (789.61) (method A).

Example 411 Preparation of Compound 411

Step 1:

To a solution of Boc-L-hydroxyproline (2.00 g, 8.65 mmol) in THF (25mL), NaH (0.795 g, 19.87 mmol) was added under a nitrogen atmosphere.The suspension was stirred at room temperature for 15 minutes, and4,6-dichloropyrimidine (2.58 g, 17.30 mmol) was added in two portions.The mixture was stirred at room temperature for one hour and 1.3equivalents of HCl (1N) were used to neutralize the reaction. pH 4.0buffer solution was added and the pH was adjusted to pH 5. Ethyl acetatewas used to extract the aqueous phase (3×25 mL) and the organic layerswere washed with brine (20 mL) and dried over MgSO₄ to yield the titledcompound as a white solid (crude yield not calculated). The crudeproduct was taken into the next step.

LC/MS rt-min (MH⁺): 1.91 (366.2) (method A).

Step 2:

A solution of the crude product from Step 1, (assumed 8.65 mmol), in THF(40 mL) and methanol (40 mL) was cooled to 0° C. TMSCN₂ 2M in hexanes(˜1.3 eq) was slowly added to the stirring solution under a nitrogenatmosphere until gas was no longer emitted from the solution. The fullyreacted solution was then concentrated in vacuo, and purification by aBiotage 40M column (eluted 20%-40% ethyl acetate in hexanes) to affordthe pure titled compound as a white foam (497 mg, 16% over steps 2a-2b).

¹H NMR: (DMSO-d₆) δ 1.34-1.38 (rotamers, 2:1, 9H), 2.25-2.29 (m, 1H),2.53-2.56 (m, 1H), 3.58-3.75 (m, 2H), 3.69 (s, 3H), 4.28-4.33 (m, 1H),5.59 (s, 1H), 7.24 (s, 1H), 8.69 (s, 1H);

LC/MS rt-min (MH⁺): 2.08 (380.14) (method A).

Step 3:

A solution of the pure product from Step 2 (472 mg, 1.83 mmol) in DCM (3mL) and TFA (5.65 mL) was stirred at room temperature for one hour. Thesolvent was removed in vacuo, the residue was suspended in 1N HCl indiethyl ether (7.33 mL), gently stirred and concentrated in vacuo. Thisprocedure was repeated, and the resulting product was placed on an oilpump overnight to yield a white solid in quantitative yield.

LC/MS rt-min (MH⁺): 0.55 (258.35) (method A).

Step 4:

The product from Step 3 (assumed quantitative yield, 1.83 mmol) wasadded to a solution of BOC-t-Butyl-L-glycine (0.424 g, 1.83 mmol) in DCM(11 mL) under a nitrogen atmosphere. This step was followed by theaddition of HOBt (0.272 g, 2.02 mmol), DIPEA (2.23 mL, 12.82 mmol), andHBTU (1.04 g, 2.75 mmol). The reaction was left to stir at roomtemperature for 15 hours. 15 mL DCM was added to the solution in orderto increase the volume, and the reaction was quenched with pH 4.00buffer solution (15 mL). The mixture was acidified to a pH of 4.5 using1N HCl, and the aqueous phase was extracted with DCM (3×20 mL). Theorganic phase was washed twice with pH 4.00 buffer solution (20 mL),saturated NaOH (25 mL), brine (20 mL), and dried over MgSO₄. Theresulting solution was concentrated in vacuo and purified by a Biotage40S column (eluted 20%-50% ethyl acetate in hexanes). This purificationafforded the pure titled compound as a white solid (454 mg, 53%).

¹H NMR: (DMSO-d₆) δ 0.94 (s, 9H), 1.25 (s, 9H), 2.21-2.27 (m, 1H),2.48-2.55 (m, 1H), 3.64 (s, 3H), 3.86-4.02 (m, 2H), 4.29-4.31 (d, J=10Hz, 1H), 4.46-4.49 (t, 1H), 5.75 (br s, 1H), 6.72-6.74 (d, NH), 7.12 (s,1H), 8.71 (s, 1H);

LC/MS rt-min (MH⁺): 2.27 (493.5) (method A).

Step 5:

LiOH (0.0141 g, 0.589 mmol) was dissolved in water (7.5 mL) by heatingand sonication. The LiOH solution was added to a solution of the purematerial from Step 4 (252 mg, 0.535 mmol) in THF (7.5 mL), and left tostir at room temperature. The reaction was complete after 3 hours. Itwas quenched with pH 4.0 buffer and acidified to a pH of approximately4.5 with 1N HCl. The aqueous phase was extracted with EtOAc (3×25 mL),and the organic phase was washed with brine (20 mL) and dried overMgSO₄. The filtered solution was concentrated in vacuo and left on avacuum line overnight. The crude product (231 mg, 95% yield) was takeninto the next step.

¹H NMR: (DMSO-d₆) δ 0.94 (s, 9H), 1.25 (s, 9H), 2.14-2.22 (m, 1H),2.50-2.54 (m, 1H), 3.84-3.876 (d, J=150 Hz, 1H), 3.97-3.99 (d, J=10 Hz,1H), 4.27-4.30 (d, J=Hz, 1H), 4.37-4.40 (t, 1H), 5.63 (br s, 1H),6.69-6.71 (d, NH), 7.12 (s, 1H), 8.71 (s, 1H), 12.56 (br s, OH);

LC/MS rt-min (MH⁺): 2.24 (479.5) (method A).

Step 6:

The pure material from Step 5 (80 mg, 0.146 mmol), and phenylboronicacid (0.0178 g, 0.146 mmol) were solvated in DMF (2 mL). The solutionwas placed under a nitrogen atmosphere and 2M aqueous Na₂CO₃ (0.146 mL,0.292 mmol) was added. Five mole percent ofTetrakis(triphenyl)phosphine)-palladium (0) was added (8.44 mg, 0.0073mmol) and the mixture was heated by microwave using the PersonalChemistry Emrys Optimizer for 50 minutes at 140° C. Palladium blackprecipitated out of the reaction upon completion. The mixture wasacidified with one equivalent of 1N HCl, and filtered through a syringe,using MeOH to extract the product. The product was purified by prep HPLC(column—4 Xterra S5 30×75 mm, solvent—70% A/30% B-30% A/70% B (wheresolvent A is 10% MeOH, 90% H₂O, 0.1% TFA and solvent B is 90% MeOH, 10%H₂O, 0.1% TFA), gradient time—15 min., hold time—1 min., flow rate—40mL/min, retention time of pure product—10.45-11.37). Fractionscontaining the desired product were neutralized with 1N NaOH and placedin the speed vacuum for approximately 4 hours. The fractions werecombined and pH 4.0 buffer (15 mL) was added. The pH was adjusted to pH4-5 using 1N HCl, and the aqueous layer was extracted with ethyl acetate(3×20 mL). The organic layer was washed with brine (15 mL), dried overMgSO₄, and concentrated in vacuo. The product was placed on an oil pumpto dry overnight, and an viscous oil was obtained (37 mg, 50%).

LC/MS rt-min (MH⁺): 2.37 (499.3) (method A).

Step 7:

The product from Example 411, Step 6 (36.7 mg, 0.061 mmol) was dissolvedin DCM (4 mL) and cyclopropanesulfonic acid (1(R)-amino-2(S)-vinyl-cyclopropanecarbonyl)-amide hydrochloride salt (Example 1,Step 8) (0.0164 g, 0.161 mmol) was added. DIPEA (0.0534 mL, 0.307 mmol),and then HATU (0.0326, 0.0858 mmol) were added under a nitrogenatmosphere. The reaction was left to stir at room temperature for 3hours. 10 mL DCM was added to the solution in order to increase thevolume, and the reaction was quenched with pH 4.00 buffer solution (10mL). The mixture was acidified to a pH of 4-5 using 1N HCl, and theaqueous phase was extracted with DCM (3×15 mL). The organic phase waswashed twice with pH 4.00 buffer solution (10 mL) and brine (10 mL), andthen dried over MgSO₄. The resulting solution was concentrated in vacuoand purified by a Biotage 12M column (eluted 20%-40% acetone inhexanes). This purification afforded the pure Compound 411 as a whitepowder (7 mg, 15%).

¹H NMR: δ 1.07-1.46 (m, 24H), 1.87-1.90 (m, 1H), 2.22-2.32 (m, 2H),2.51-2.55 (m, 1H), 2.92-2.97 (m, 1H), 4.04-4.07 (d, J=15 Hz, 1H),4.20-4.22 (d, J=10 Hz, 1H), 4.36-4.38 (d, J=10 Hz, 1H), 4.47-4.50 (t,1H), 5.12-5.14 (d, J=10 Hz, 1H), 5.29-5.33 (d, J=20 Hz, 1H), 5.73-5.82(m, 2H), 6.59-6.60 (d, NH), 7.29 (s, 1H), 7.50-7.51 (m, 3H), 8.03-8.05(m, 2H), 8.81 (s, 1H);

LC/MS rt-min (MH⁺): 2.50 (711.4) (method A).

Example 412 Preparation of Compound 412

Step 1:

The product from Example 411, Step 5 (80 mg, 0.146 mmol) was solvated inDMF (2 mL), and 2-thiopheneboronic acid (0.028 g, 0.219 mmol) was addedto the solution. The reaction was placed under a nitrogen atmosphere and2M aqueous Na₂CO₃ (0.146 mL, 0.292 mmol), and 5 mole percent ofTetrakis(triphenyl)phosphine)-palladium (0) were added (8.44 mg, 0.0073mmol). The reaction was heated by microwave using the Personal ChemistryEmrys Optimizer for 30 minutes at 150° C. Palladium black precipitatedout of the reaction upon completion. The mixture was acidified with oneequivalent of 1N HCl, and filtered through a syringe, using MeOH toextract the product. The product was purified by prep HPLC (column—4Xterra S5 30×75 mm, solvent—70% A/30% B-30% A/70% B (where solvent A is10% MeOH, 90% H₂O, 0.1% TFA and solvent B is 90% MeOH, 10% H₂O, 0.1%TFA), gradient time—15 min., hold time—1 min., flow rate—40 mL/min,retention time of pure product—10.45-11.37). The fractions containingthe desired product were neutralized with 1N NaOH and placed in thespeed vacuum for approximately 4 hours. The fractions were thencombined, and pH 4.0 buffer (15 mL) was added. The pH was adjusted to pH4-5 using 1N HCl and the aqueous layer was extracted with ethyl acetate(3×20 mL). The organic layer was washed with brine (15 mL), dried overMgSO₄, and concentrated in vacuo. The product was placed on an oil pumpto dry overnight, and an oily liquid was obtained (37 mg, 50%).

LC/MS rt-min (MH⁺): 2.37 (499.3) (method A).

Step 2:

The product from Example 412, Step 1 (39 mg, 0.0773 mmol) was dissolvedin DCM (4 mL) and cyclopropanesulfonic acid (1(R)-amino-2(S)-vinyl-cyclopropanecarbonyl)-amide hydrochloride salt (Example 1,Step 8) (0.0206 g, 0.0773 mmol) was added. DIPEA (0.015 mL, 0.387 mmol),and then HATU (0.0411 g, 0.108 mmol) were added under a nitrogenatmosphere. The reaction was left to stir at room temperature for 15hours. 10 mL DCM was added to the solution in order to increase thevolume, and the reaction was quenched with pH 4.00 buffer solution (10mL). The mixture was acidified to a pH of 4-5 using 1N HCl and theaqueous phase was extracted with DCM (3×15 mL). The organic phase waswashed twice with pH 4.00 buffer solution (10 mL) and brine (10 mL), andthen dried over MgSO₄. The resulting solution was concentrated in vacuoand purified by a Biotage 12M column (eluted 20%-50% acetone inhexanes). This purification afforded the pure Compound 412 as a whitepowder (4 mg, 7%).

¹H NMR: δ 1.04-1.29 (m, 24H), 1.45-1.47 (m, 1H), 1.89-1.91 (m, 1H),2.26-2.31 (m, 2H), 2.51-2.53 (m, 1H), 2.92-2.95 (m, 1H), 4.05-4.07 (d,J=10 Hz, 1H), 4.22-4.24 (d, J=10 Hz, 1H), 4.38-4.40 (d, J=10 Hz, 1H),4.48-4.52 (m, 1H), 5.15-5.17 (d, J=10 Hz, 1H), 5.32-5.36 (d, J=20 Hz,1H), 5.76-5.83 (m, 2H), 6.65-6.67 (d, NH), 7.19-7.21 (m, 2H), 7.66-7.67(d, J=5 Hz, 1H), 7.86-7.87 (d, J=5 Hz, 1H), 8.69 (s, 1H);

LC/MS rt-min (MH⁺): 2.45 (739.4) (method A).

Example 413 Preparation of Compound 413

Step 1:

2-Bromo-6-chloropyridine (3.0 g, 15.55 mmol) and phenylboronic acid(1.896 g, 15.55 mmol) were solvated in a mixture of EtOH, toluene andwater (2:1:1; 120 mL). Aqueous 1M Na₂CO₃ (15.55 mL, 31.10 mmol) and 5mole percent of Tetrakis(triphenyl)phosphine)-palladium (0) (0.896 g,0.775 mmol) were added under a nitrogen atmosphere. The reaction wasrefluxed at 90° C. for one hour. Water (20 mL) was added to quench thereaction, and the aqueous layer was extracted with diethyl ether (4×25mL). The organic layer was then washed with brine, dried over MgSO₄, andconcentrated in vacuo. The crude mixture was purified by a Biotage 40Scolumn (eluted 2%-10% ethyl acetate in hexanes), to afford the puretitled compound (1.45 g, 74%).

¹H NMR: δ 7.24-7.26 (m, 1H), 7.42-7.48 (m, 3H), 7.63-7.70 (m, 2H),7.98-8.00 (m, 2H);

LC/MS rt-min (MH⁺): 2.04 (190.18) (method A)

Step 2:

TFA (20 mL) was added to the pure solid obtained in Step 1 (3.27 g,17.24 mmol). A 30% solution of H₂O₂ (5.55 mL, 48.9 mmol) was slowlyadded dropwise to the stirring solution under a nitrogen atmosphere. Thereaction was heated to reflux at 100° C. for 3 hours, and 0.5 additionalequivalents of H₂O₂ (2.27 mL, 25 mmol) were added to the solution. Thereaction continued to stir at 100° C. for 2 hours. The flask was allowedto cool to room temperature before the solution was concentrated invacuo to approximately half of the original volume. Water (40 mL) wasadded to quench the reaction and the aqueous layer was extracted withethyl acetate (5×30 mL). The organic layer was washed once with brine(20 mL), dried over MgSO₄, and concentrated in vacuo. The crude productwas purified by Biotate 40M column (eluted 10%-75% ethyl acetate inhexanes) to yield a pale yellow liquid (0.81 g, 23%). Pure startingmaterial was also recovered for future use (1.895 g, 58%).

¹H NMR: δ 7.40-7.42 (t, 1H), 7.49-7.50 (m, 3H), 7.59-7.61 (d, J=10 Hz,1H), 7.77-7.82 (m, 3H);

LC/MS rt-min (MH⁺): 1.12 (206.37) (method A).

Step 3:

The purified product from Step 2 (0.81 g, 3.94 mmol) was added to asolution of SOCl₂ (25 mL) and stirred at 60° C. for 2 hours. Thetemperature was then increased to 80° C. in order to force the reactionto completion, and it was heated for an additional 30 min. The solutionwas concentrated in vacuo and quenched carefully with ice. The pH wasadjusted to a pH of 4-5 using 10N NaOH, keeping the flask in an icebath. The aqueous phase was extracted with ether (4×25 mL), and theorganic layer was washed with brine (20 mL), dried over MgSO₄, andconcentrated in vacuo. The crude, yellow liquid was purified by Biotage40S column (eluted 2%-10% ethyl acetate in hexanes). A slightly yellow,viscous liquid was obtained (538 mg, 61%).

¹H NMR: δ 7.52-7.55 (m, 3H), 7.73 (s, 1H), 8.10-8.11 (m, 2H), 8.17 (s,1H);

LC/MS rt-min (MH⁺): 2.62 (225.33) (method A).

Step 4:

To a solution of Boc-L-hydroxyproline (555 mg, 2.40 mmol) in DMSO (4mL), KO^(t)Bu (0.619 g, 5.52 mmol) was added under a nitrogenatmosphere. The suspension was stirred at room temperature for 45 min.,and 2,4-dichloro-6-phenyl-pyridine (purified in Step 3). (538 mg, 2.40mmol) was added in two portions. The mixture was stirred at roomtemperature for two hours and 1.3 equivalents of HCl (1N) were used toneutralize the reaction. Buffer solution of pH 4.0 was added, and the pHwas adjusted to pH 4-5. Ethyl acetate was used to extract the aqueousphase, (3×25 mL) and the organic phase was washed with brine (20 mL) anddried over MgSO₄ to yield the titled compound as a white solid (crudeyield=962 mg). The crude product was taken into the next step.

LC/MS rt-min (MH⁺): 2.55 (419.27) (method A).

Step 5:

A solution of the crude product from Step 4 (assumed 2.4 mmol) in THF(10 mL) and methanol (10 mL) was cooled to 0° C. TMSCN₂ 2M in hexanes(˜1.3 eq) was slowly added to the stirring solution under a nitrogenatmosphere until gas was no longer emitted from the solution. The fullyreacted solution was then concentrated in vacuo, and purification by aBiotage 25S column (eluted 10%-50% ethyl acetate in hexanes) affordedthe pure titled compound as a white foam (503 mg, 48% over steps4d-4-e).

¹H NMR: (DMSO-d₆) δ 1.35-1.38 (rotamers, δ: 2, 9H), 2.21-2.26 (m, 1H),2.50-2.58 (m, 1H), 3.67-3.70 (m, 5H), 4.26-4.34 (m, 1H), 5.35 (br s,1H), 7.14-7.15 (m, 1H), 7.47-7.55 (m, 4H), 8.05-8.09 (m, 2H);

LC/MS rt-min (MH⁺): 2.69 (455.52) (method A).

Step 6:

A solution of the pure product from Step 5 (503 mg, 1.16 mmol) in DCM(2.5 mL) and TFA (3.58 mL) was stirred at room temperature for one hour.The solvent was removed in vacuo. and the residue was suspended in 1NHCl in diethyl ether (6 mL), gently stirred, and concentrated in vacuo.This procedure was repeated and the resulting product was placed on anoil pump to yield a white solid in quantitative yield. The crude productwas carried into the next step.

Step 7:

The product from Step 6 (assumed quantitative yield, 1.83 mmol) wasadded to a solution of BOC-t-Butyl-L-glycine (0.424 g, 1.83 mmol) in DCM(11 mL) under a nitrogen atmosphere. This step was followed by theaddition of HOBt (0.272 g, 2.02 mmol), DIPEA (2.23 mL, 12.82 mmol), andHBTU (1.04 g, 2.75 mmol). The reaction was left to stir at roomtemperature for 15 hours. 15 mL DCM was added to the solution in orderto increase the volume, and the reaction was quenched with pH 4.00buffer solution (15 mL). The mixture was acidified to a pH of 4.5 using1N HCl, and the aqueous phase was extracted with DCM (3×20 mL). Theorganic phase was washed twice with pH 4.00 buffer solution (20 mL),saturated NaOH (25 mL), brine (20 mL), and dried over MgSO₄. Theresulting solution was concentrated in vacuo and purified by a Biotage40S column (eluted 20%-50% ethyl acetate in hexanes). This purificationafforded the pure titled compound as a white solid (454 mg, 53%).

¹H NMR: (DMSO-d₆) δ 0.96 (s, 9H), 1.19 (s, 9H), 2.17-2.29 (m, 1H),2.48-2.58 (m, 1H), 3.65 (s, 3H), 3.81-3.89 (m, 1H), 4.05-4.08 (m, 1H),4.21-4.26 (m, 1H), 4.44-4.50 (t, 1H), 5.45 (br s, 1H), 6.72-6.75 (d,J=15 Hz, 1H), 7.09 (s, 1H), 7.46-7.51 (m, 4H), 8.02-8.06 (m, 2H);

LC/MS rt-min (MH⁺): 2.27 (493.5) (method A).

Step 8:

LiOH (0.0233 g, 0.973 mmol) was dissolved in water (10 mL) by heatingand sonication. The LiOH solution was added to a solution of the purematerial from Step 7 (483 mg, 0.885 mmol) in THF (10 mL), and themixture immediately turned a pale peach color. The reaction was left tostir at room temperature for 1 hour and was acidified with 1N HCl (0.973mL, 0.974 mmol). The reaction was quenched with pH 4.00 buffer solutionand the pH was adjusted to a pH between 4 and 5. The aqueous layer wasextracted with EtOAc (3×20 mL), washed with brine (15 mL) and dried overMgSO₄. The filtered solution was concentrated in vacuo and left on avacuum line overnight. The crude product (480 mg, >95% yield) was takeninto the next step.

¹H NMR: (DMSO-d₆) δ 0.94 (s, 9H), 1.20 (s, 9H), 1.32-1.34 (m, 1H),2.12-2.4 (m, 1H), 2.51-2.55 (m, 1H), 3.81-3.83 (d, J=10 Hz, 1H),4.21-4.23 (d, J=10 Hz, 1H), 4.32-4.35 (t, 1H), 5.4 (br s, 1H), 6.63-6.65(d, NH), 7.09 (s, 1H), 7.47-7.49 (m, 4H), 8.03-8.06 (m, 2H), 12.56 (s,1H).

Step 9:

The dipeptide from Step 8 (100 mg, 0.188 mmol) and 2-thiopheneboronicacid (0.0481 g, 0.376 mmol) were solvated in DMF (2 mL). The solutionwas placed under a nitrogen atmosphere and 2M aqueous KF (0.282 mL,0.376 mmol) was added. Five mole percent ofTetrakis(triphenyl)phosphine)-palladium (0) was added (0.011 mg, 0.0094mmol) and the mixture was heated by microwave using the PersonalChemistry Emrys Optimizer for 30 minutes at 150° C. Palladium blackprecipitated out of the reaction upon completion. The mixture wasacidified with one equivalent of 1N HCl and filtered through a syringe,using MeOH to extract the product. The product was purified by prep HPLC(column—4 Xterra S5 5 um 30×75 mm, solvent—85% A/15% B-10% A/90% B(where solvent A is 10% MeOH, 90% H₂O, 0.1% TFA and solvent B is 90%MeOH, 10% H₂O, 0.1% TFA), gradient time—15 min., hold time—1 min., flowrate—40 mL/min, retention time of pure product—16.23). Fractionscontaining the desired product were neutralized with 1N NaOH and placedin the speed vac for approximately 2 hours. The fractions were combined,and pH 4.0 buffer (15 mL) was added. The pH was adjusted to pH 4-5 using1N HCl, and the aqueous layer was extracted with ethyl acetate (3×20mL). The organic layer was washed with brine (15 mL), dried over MgSO₄,and concentrated in vacuo. The product was placed on an oil pump to dryovernight, and a pale yellow oil was obtained (44 mg, 40%).

LC/MS rt-min (MH⁺): 2.7 (580.54) (method A).

Step 10:

The product from Example 413, Step 9 (43 mg, 0.0742 mmol) was dissolvedin DCM (2 mL) and cyclopropanesulfonic acid (1(R)-amino-2(S)-vinyl-cyclopropanecarbonyl)-amide hydrochloride salt (Example 1,Step 8) (0.0198 g, 0.0742 mmol) was added. DIPEA (0.0646 mL, 0.371mmol), and then HATU (0.039 g, 0.0104 mmol) were added under a nitrogenatmosphere. The reaction was left to stir at room temperature for 4.5hours. 10 mL of DCM was added to the solution in order to increase thevolume, and the reaction was quenched with pH 4.00 buffer solution (10mL). The mixture was acidified to a pH of 4-5 using 1N HCl, and theaqueous phase was extracted with DCM (3×15 mL). The organic phase waswashed twice with pH 4.00 buffer solution (10 mL) and brine (10 mL), andthen dried over MgSO₄. The resulting solution was concentrated in vacuoand purified by a Biotage 12S column (eluted 10%-50% acetone inhexanes). The compound was isolated and re-purified by prep-HPLC(column—YMC ODS-A 20×50 mm s5, solvent—60% A/40% B-10% A/90% B (wheresolvent A is 10% MeOH, 90% H₂O, 0.1% TFA and solvent B is 90% MeOH, 10%H₂O, 0.1% TFA), gradient time—8 min., hold time—2 min., flow rate—25mL/min, retention time of pure product—8.702). This purificationafforded the pure Compound 413 as a pale orange oil (22.3 mg, 38%).

¹H NMR: δ 1.02-1.29 (m, 24H), 1.42-1.45 (m, 1H), 1.87-1.89 (m, 1H),2.24-2.30 (m, 2H), 2.53-2.57 (m, 1H), 2.92-2.97 (m, 1H), 4.06-4.08 (d,J=10 Hz, 1H), 4.25 (s, 1H), 4.31-4.33 (d, J=10 Hz, 1H), 4.45-4.49 (t,1H), 5.12-5.14 (d, J=10 Hz, 1H), 5.29-5.32 (d, J=15 Hz, 1H), 5.46 (br s,1H), 5.73-5.80 (m, 1H), 7.13-7.15 (m, 1H), 7.29-7.30 (d, J=5 Hz, 2H),7.42-7.53 (m, 4H), 7.75-7.56 (d, J=5 Hz, 1H), 8.07-8.09 (d, J=10 Hz,2H).

LC/MS rt-min (MH⁺): 2.79 (792.72) (method A).

Example 414 Preparation of Compound 414

Step 1:

The pure material from Example 413, Step 8 (100 mg, 0.188 mmol), and4-methoxyphenylboronic acid (0.0429 g, 0.282 mmol) were solvated in DMF(2.5 mL). The solution was placed under a nitrogen atmosphere and 2Maqueous Na₂CO₃ (0.188 mL, 0.376 mmol) was added. Five mole percent ofTetrakis(triphenyl)phosphine)-palladium (0) was added (0.011 mg, 0.0094mmol), and the mixture was heated by microwave using the PersonalChemistry Emrys Optimizer for 30 minutes at 150° C. Palladium blackprecipitated out of the reaction upon completion. The mixture wasacidified with one equivalent of 1N HCl and filtered through a syringe,using MeOH to extract product. The product was purified by prep HPLC(column—Xterra MS C18 5 um 30×50 mm, solvent—90% A/10% B-10% A/90% B(where solvent A is 10% MeOH, 90% H₂O, 0.1% TFA and solvent B is 90%MeOH, 10% H₂O, 0.1% TFA), gradient time—15 min., hold time—1 min., flowrate—45 mL/min, retention time of pure product—13.72). Fractionscontaining the desired product were neutralized with 1N NaOH and placedin the speed vac for approximately 2 hours. The fractions were combined,and pH 4.0 buffer (15 mL) was added. The pH was adjusted to pH 4-5 using1N HCl, and the aqueous layer was extracted with ethyl acetate (3×20mL). The organic layer was washed with brine (15 mL), dried over MgSO₄,and concentrated in vacuo. The product was placed on an oil pump to dryovernight, and a viscous oil was obtained (44 mg, 50%).

LC/MS rt-min (MH⁺): 2.23 (604.61) (method A).

Step 2:

The product from Example 414, Step 1 (43 mg, 0.0712 mmol) was dissolvedin DCM (2 mL) and cyclopropanesulfonic acid (1(R)-amino-2(S)-vinyl-cyclopropanecarbonyl)-amide hydrochloride salt (Example 1,Step 8) (0.01899 g, 0.0712 mmol) was added. DIPEA (0.062 mL, 0.356mmol), and then HATU (0.038 g, 0.0997 mmol) were added under a nitrogenatmosphere. The reaction was left to stir at room temperature for 4.5hours. 10 mL DCM was added to the solution in order to increase thevolume, and the reaction was quenched with pH 4.00 buffer solution (10mL). The mixture was acidified to a pH of 4-5 using 1N HCl, and theaqueous phase was extracted with DCM (3×15 mL). The organic phase waswashed twice with pH 4.00 buffer solution (10 mL) and brine (10 mL), andthen dried over MgSO₄. The resulting solution was concentrated in vacuoand purified by a Biotage 12S column (eluted 20%-50% acetone inhexanes). The compound was isolated and re-purified by prep-HPLC(column—YMC ODS-A 20×50 mm s5, solvent—60% A/40% B-10% A/90% B (wheresolvent A is 10% MeOH, 90% H₂O, 0.1% TFA and solvent B is 90% MeOH, 10%H₂O, 0.1% TFA), gradient time—10 min., hold time—2 min., flow rate—25mL/min, retention time of pure product—7.43-8.24). This purificationafforded the pure Compound 414 as a pale orange oil (19.4 mg, 34%).

¹H NMR: δ 1.02-1.23 (m, 24H), 1.29-1.44 (m, 1H), 1.89-1.95 (m, 1H),2.24-2.30 (q, 1H), 2.32-2.40 (m, 1H), 2.59-2.62 (m, 1H), 3.90 (s, 3H),4.10-4.12 (d, J=10 Hz, 1H), 4.21 (s, 1H), 4.42-4.56 (m, 2H), 5.12-5.14(d, J=10 Hz, 1H), 5.28-5.31 (d, J=Hz, 1H), 5.61 (br s, 1H), 5.72-5.80(m, 1H), 7.14-7.16 (d, J=10 Hz, 2H), 7.52-7.54 (d, J=10 Hz, 2H),7.61-7.62 (m, 2H), 7.95-7.98 (m, 4H);

LC/MS rt-min (MH⁺): 2.35 (816.76) (method A).

Example 415 Preparation of Compound 415

Step 1:

The dipeptide from Example 413, Step 8 (174 mg, 0.327 mmol) andphenylboronic acid (0.06 g, 0.491 mmol) were solvated in DMF (4 mL). Thesolution was placed under a nitrogen atmosphere and 2M aqueous NaCO₃(0.33 mL, 0.654 mmol) was added. Five mole percent ofTetrakis(triphenyl)phosphine)-palladium (0) was added (0.019 mg, 0.0164mmol) and the mixture was heated by microwave using the PersonalChemistry Emrys Optimizer for 30 minutes at 150° C. Palladium blackprecipitated out of the reaction upon completion. The mixture wasacidified with one equivalent of 1N HCl and filtered through a syringe,using MeOH to extract the product. The product was purified by prep HPLC(column—5 Xterra c-18 5 um 30×100 mm, solvent—80% A/20% B-0% A/100% B(where solvent A is 10% MeOH, 90% H₂O, 0.1% TFA and solvent B is 90%MeOH, 10% H₂O, 0.1% TFA), gradient time—20 min., hold time—1 min., flowrate—40 mL/min, retention time of pure product—11.28-11.72). Fractionscontaining the desired product were neutralized with 1N NaOH and placedin the speed vac for approximately 2 hours. The fractions were combined,and pH 4.0 buffer (15 mL) was added. The pH was adjusted to pH 4-using1N HCl, and the aqueous layer was extracted with ethyl acetate (4×15mL). The organic layer was washed with brine (15 mL), dried over MgSO₄,and concentrated in vacuo. The product was placed on an oil pump to dryovernight. (31.5 mg, 17%).

LC/MS rt-min (MH⁺): 2.54 (574.37) (method A).

Step 2:

The product from Example 415, Step 1 (31.5 mg, 0.055 mmol) was dissolvedin DCM (3 mL) and cyclopropanesulfonic acid (1(R)-amino-2(S)-vinyl-cyclopropanecarbonyl)-amide hydrochloride salt (Example 1,Step 8) (0.0147 g, 0.055 mmol) was added. DIPEA (0.048 mL, 0.275 mmol),and then HATU (0.029 g, 0.077 mmol) were added under a nitrogenatmosphere. The reaction was left to stir at room temperature for 3.5hours and an additional 0.3 equivalents of cyclopropanesulfonic acid(1(R)-amino-2 (S)-vinyl-cyclopropanecarbonyl)-amide hydrochloride saltwere added. The reaction was left to stir for 8 more hours. 10 mL of DCMwas added to the solution in order to increase the volume, and thereaction was quenched with pH 4.00 buffer solution (10 mL). The mixturewas acidified to a pH of 4-5 using 1N HCl, and the aqueous phase wasextracted with DCM (4×10 mL). The organic phase was washed twice with pH4.00 buffer solution (10 mL) and brine (10 mL), and then dried overMgSO₄. The resulting solution was concentrated in vacuo and purified byprep HPLC (column—YMC ODS-A 30×50 mm, solvent—80% A/20% B-10% A/90% B(where solvent A is 10% MeOH, 90% H₂O, 0.1% TFA and solvent B is 90%MeOH, 10% H₂O, 0.1% TFA), gradient time—20 min., hold time—3 min., flowrate—30 mL/min, retention time of pure product—19.6). This purificationdid not afford a pure compound so it was re-purified by a Biotage 12 Scolumn (eluted 10%-50% acetone in hexanes). This purification affordedthe pure titled compound as a pale orange oil (22.3 mg, 38%).

¹H NMR: δ 1.02-1.45 (m, 24H), 1.85-1.86 (m, 1H), 2.03-2.11 (m, 2H),2.42-2.46 (m, 1H), 2.77-2.85 (m, 1H), 4.10-4.12 (m, 1H), 4.25 (s, 1H),4.31-4.33 (d, J=10 Hz, 1H), 4.49-4.54 (m, 1H), 5.03-5.05 (d, J=10 Hz,1H), 5.20-5.24 (d, J=20 Hz, 1H), 5.44 (br s, 1H), 5.82-5.94 (m, 1H),7.33 (s, 2H), 7.43-7.50 (m, 6H), 8.09-8.10 (d, J=Hz, 4H);

LC/MS rt-min (MH⁺): 2.37 (786.37) (method A).

Section J

In section J the LC/MS method utilized was the following:

Columns:

-   -   Method A: YMC ODS-A C18 S7 (4.6×33 mm)    -   Method B: YMC Xterra ODS S7 (3.0×50 mm)    -   Method C: Xterra ms C18 (4.6×33 mm)    -   Method D: YMC ODS-A C18 S3 (4.6×33 mm)        Gradient: 100% solvent A/0% solvent B to 0% solvent A/100%        solvent B        Gradient time: 3 min.        Hold Time: 1 min.        Flow Rate: 5 mL/min.        Detector Wavelength: 220 nm.        Solvents: Solvent A: 10% MeOH/90% water/0.1% TFA. Solvent B: 90%        MeOH/10% water/0.1% TFA.

Example 420 Preparation of Compound 420

Step 1:

To a solution of PPh₃ (16.8 g, 63.9 mmol) in THF (150 mL) was addeddropwise DEAD (10.1 mL, 63.9 mmol) and the solution was stirred for 30min at room temperature. A solution of Boc-cis-(L)-Hyp-OMe (10.5 g, 42.6mmol) in THF (50 mL) was added slowly, followed by 5-bromo-pyridin-3-ol(prepared according to F. E. Ziegler et al., J. Am. Chem. Soc., (1973),95, 7458) (8.90 g, 51.1 mmol) portion wise. The resulting solution wasstirred at 45° C. for 18 hrs. The solution was concentrated in vacuo,and the residue partitioned between diethyl ether (200 mL) and 1N NaOH(50 mL). The organic phase was dried (MgSO₄) and the ethereal solutionwas after filtration concentrated to half its volume. A precipitateformed that was removed by filtration. The filtrated was concentrated invacuo and purified by a Biotage 65 M column (eluted with hexanes-EtOAc2:1, 3:2, 1:1) to provide the title compound (11.9 g, 69%) as a red oil.

¹H NMR: (CDCl₃) δ 1.42, 1.45 (s, 9H (rotamers)), 2.25-2.30 (m, 1H),2.49-2.58 (m, 1H), 3.75, 3.81 (s, 3H (rotamers)), 3.66-3.81 (m, 2H(hidden)), 4.42, 4.50 (t, J=8 Hz, 1H (rotamers)), 4.92 (m, 1H), 7.36 (s,1H), 8.20 (s, 1H), 8.32 (s, 1H).

LC/MS rt-min (MH⁺): 2.26 (401, 403) (method B).

Step 2:

The product of Step 1 (2.34 g, 5.83 mmol) was dissolved in DCM (20 mL)and TFA (18 mL, 0.23 mole) and stirred at room temperature for 2 hrs.The volatiles were removed in vacuo and the residue partitioned betweenEtOAc and water. The organic phase was extracted with 1N HCl (25 mL) andthe combined aqueous extracts were neutralized with satd. NaHCO₃ (40mL). The aqueous phase was extracted with EtOAc (twice) and the combinedorganic extracts washed with brine and dried (MgSO₄) to give the titlecompound (1.02 g, 58%, free base) as a colorless oil.

¹H NMR: (DMSO-d₆) δ 2.12-2.20 (m, 2H), 2.94 (d, J=12 Hz, 1H), 3.19 (dd,J=4.5, 12 Hz, 1H), 3.64 (s, 3H), 3.90 (t, J=8 Hz, 1H), 5.07 (m, 1H),7.69 (s, 1H), 8.26 (s, 1H), 8.29 (s, 1H).

LC/MS rt-min (MH⁺): 0.78 (301, 302) (method B).

Step 3:

To a suspension of the product of Step 2 (1.02 g, 3.39 mmol),N—BOC-L-tert-leucine (862 mg, 3.73 mmol), and HOBt (458 mg, 3.39 mmol)in DCM (15 mL) were added DIPEA (2.36 mL, 13.6 mmol) followed by HBTU(1.61 g, 4.24 mmol). The resulting solution was stirred at roomtemperature for 15 hrs and quenched with DCM and buffer pH 4 and some 1NHCl to adjust the pH to 4-5. The organic phase was washed with buffer pH4, satd. NaHCO₃ (twice), brine, and dried (MgSO₄). Purification using aBiotage 40 M column (eluted hexane-EtOAc 3:2, 1:1) afforded the titlecompound (1.48 g, 85%) as a white solid.

¹H NMR: (DMSO-d₆) δ 0.94 (s, 9H), 1.17 (s, 9H), 2.16-2.21 (m, 1H), 2.50(m, (hidden), 1H), 3.64 (s, 3H), 3.82 (d, J=12 Hz, 1H), 4.06 (d, J=9.5Hz, 1H), 4.16 (d, J=12 Hz, 1H), 4.45 (dd, J=8, 9.5 Hz, 1H), 5.26 (m,1H), 6.71 (d, J=9 Hz, NH), 7.75 (s, 1H), 8.28 (s, 1H), 8.32 (s, 1H).

LC/MS rt-min (MH⁺): 2.35 (514, 516) (method B).

Step 4:

To a mixture of the product of Step 3 (98 mg, 0.19 mmol), Pd(PPh₃)₄ (6.6mg, 0.00573 mmol), 2M aqueous Na₂CO₃ (0.191 mL, 0.381 mmol) in toluene(2 mL) was added a solution of phenyl boronic acid (29 mg, 0.24 mmol) inmethanol (0.1 mL). The solution was heated at 85° C. for 6 h undernitrogen. After cooling to room temperature the mixture was quenchedwith buffer pH 4 and extracted with EtOAc (2×10 mL), and dried (Na₂SO₄).Purification using a Biotage 12 M column (eluted hexane-EtOAc 2:3)afforded the title compound (72 mg, 74%) as a colorless oil.

¹H NMR: (methanol-d₄) δ 1.05 (s, 9H), 1.31 (s, 9H), 2.29-2.35 (m, 1H),2.68-2.72 (m, 1H), 3.77 (s, 3H), 4.00 (d, J=12 Hz, 1H), 4.24 (d, J=9.5Hz, 1H), 4.41 (d, J=12 Hz, 1H), 4.67 (dd, J=7.5, 10 Hz, 1H), 5.36 (m,1H), 6.51 (d, J=9.5 Hz, NH), 7.79-7.85 (m, 3H), 8.39 (s, 1H), 8.59 (s,1H), 8.67-8.68 (m, 2H).

LC/MS rt-min (MH⁺): 2.16 (512) (method B).

Step 5:

To a solution of the product of Step 4 (150 mg, 0.293 mmol) in THF (2mL) and methanol (2 mL) was added LiOH (14 mg, 0.59 mmol) in water (2mL). The mixture was stirred for 2 h at room temperature and quenchedwith 1N HCl until neutral pH. The organic volatiles were removed invacuo and to the residue was added buffer pH 4. The product wasextracted into EtOAc (3×10 mL), washed with brine/buffer pH 4 and dried(MgSO₄) to yield the title compound in quantitative yield as a whitesolid after trituration from pentane.

¹H NMR: (methanol-d₄) δ 1.05 (s, 9H), 1.32 (s, 9H), 2.32-2.38 (m, 1H),2.69-2.74 (m, 1H), 3.99 (dd, J=3, 12 Hz, 1H), 4.25 (s, 1H), 4.40 (d,J=12 Hz, 1H), 4.64 (dd, J=7.5, 9.5 Hz, 1H), 5.36 (m, 1H), 7.85-7.87 (m,3H), 8.40 (s, 1H), 8.60 (s, 1H), 8.69-8.70 (m, 2H).

LC/MS rt-min (MH⁺): 2.03 (499) (method B).

Step 6:

To a suspension of the product of Step 5 (93 mg, 0.19 mmol), and theproduct from Example 1, Step 8 (50 mg, 0.19 mmol) in DCM (2 mL) wasadded DIPEA (0.163 mL, 0.935 mmol), followed by HATU (92 mg, 0.243mmol). The resulting mixture was stirred at room temperature for 18 hrsand quenched with DCM and buffer pH 4 and some 1N HCl to adjust the pHto 4-5. The layers were separated and the aqueous phase extracted withDCM (10 mL) and dried (Na₂SO₄). Purification using a Biotage 12 M column(eluted gradient hexane-acetone 20-60%) afforded the title compound (65mg, 49%) as a white powder.

¹H NMR: (DMSO-d₆) δ 0.95 (s, 9H), 1.03-1.05 (m, 2H), 1.08-1.10 (m, 2H),1.26 (s, 9H), 1.36-1.39 (m, 1H), 1.69-1.72 (m, 1H), 2.09-2.21 (m, 2H),2.41-2.45 (m, 1H), 2.92-2.94 (m, 1H), 3.91 (d, J=11.5 Hz, 1H), 4.08 (d,J=7.5 Hz, 1H), 4.16 (d, J=11.5 Hz, 1H), 4.36 (t, J=8.5 Hz, 1H), 5.10 (d,J=10 Hz, 1H), 5.23 (d, J=17.5 Hz, 1H), 5.38 (m, 1H), 5.59-5.67 (m, 1H),6.54 (s, NH), 7.41-7.46 (m, 1H), 7.50-7.53 (m, 2H), 7.65 (s, 1H),7.74-7.75 (m, 2H), 8.28 (s, 1H), 8.52 (s, 1H), 8.92 (s, NH).

LC/MS rt-min (MH⁺): 2.08 (710) (method B).

Example 421 Preparation of Compound 421

Step 1:

To a cold (−5° C.) 48% aqueous HBr solution (35 mL) was added4-chloro-pyridin-2-ylamine (4.18 g, 32.5 mmol; prepared according to K.S. Gudmundsson et al; Synth. Commun. (1997), 27, 861), followed by theslow addition of bromine (6.7 mL, 0.13 mole). After 20 min sodiumnitrite (8.63 g, 0.125 mole) in water (40 mL) was added at the sametemperature and stirring was continued for 30 min. The reaction wasquenched under ice cooling with 10 N NaOH (ca 40 mL) to alkaline pH. Theproduct was extracted into EtOAc (2×100 mL) and dried (Na₂SO₄). Thecrude material was purified using a Biotage 40 M column (eluted with 10%EtOAc in hexanes) to afford the title compound (2.50 g, 40%) as acolorless oil that solidified upon standing.

¹H NMR: (DMSO-d₆) δ 7.64 (dd, J=1.5, 5.5 Hz, 1H), 7.94 (d, J=1.5 Hz,1H), 8.40 (d, J=5.5 Hz, 1H);

LC/MS rt-min (MH⁺): 1.65 (192, 194, 196) (method B).

Step 2:

To a solution of N—BOC-trans-L-Hyp-OH (3.22 g, 13.9 mmol) in DMSO (30mL) was added potassium tert.butoxide (3.90 g, 34.8 mmol) in portions atroom temperature under a nitrogen atmosphere. After 1.5 h the product ofStep 1 was added in DMSO (5 mL). The mixture was stirred overnight andquenched with water (150 mL). The solution was washed with EtOAc (100mL). The aqueous phase was acidified to pH 4 with 1N HCl. The crudecarboxylic acid was extracted into EtOAc (thrice) and dried (MgSO₄). Theresidue, as a brown solid, was suspended in THF (20 mL) and MeOH (20 mL)and cooled to 0° C. A solution of trimethylsilyl diazomethane (2 M inhexane, 12 mL) was dropwise added, and the solution concentrated invacuo after 15 min. Purification using a Biotage 40 M column (elutedwith hexanes-EtOAc 2:1, 1:1) gave the title compound (3.47 g, 62%) as anoff-white powder.

¹H NMR: (DMSO-d₆) δ 1.34, 1.38 (s, 9H (rotamers)), 2.24-2.29 (m, 1H),2.43-2.50 (m, 1H), 3.57 (d, J=12.5 Hz, 1H), 3.64-3.68 (m, 1H), 3.66,3.69 (s, 3H (rotamers)), 4.27-4.34 (m, 1H), 5.21 (m, 1H), 7.06 (d, J=6Hz, 1H), 7.28 (s, 1H), 8.20 (d, J=6 Hz, 1H);

LC/MS rt-min (MH⁺): 2.59 (401, 403) (method B).

Step 3:

The product of Step 2 (2.00 g, 4.98 mmol) was suspended in 4N HCl indioxane (10 mL) and 1N HCl in diethyl ether (40 mL) and stirredovernight at room temperature. The resulting suspension was concentratedin vacuo and the residue triturated from pentane to give the titlecompound in quantitative yield as a white powder.

LC/MS rt-min (MH⁺): 0.24 (301, 303) (method B).

Step 4:

To a suspension of the product of Step 3 (assumed 4.98 mmol),N-cyclopentyloxycarbonyl-L-tert-leucine (1.24 g, 5.10 mmol), and HOBt(673 mg, 4.98 mmol) in DCM (25 mL) was added DIPEA (4.34 mL, 24.9 mmol)followed by HBTU (2.36 g, 6.23 mmol). The resulting solution was stirredat room temperature for 15 hrs and quenched with DCM and buffer pH 4 and10 mL 1N HCl to adjust the pH to 4. The aqueous phase was extracted withDCM (twice). The combined organic extracts were washed with satd. NaHCO₃(twice), brine, and dried (MgSO₄). Purification using a Biotage 40 Mcolumn (eluted hexane-EtOAc 3:2, 1:1) afforded the title compound (2.09g, 80%) as a white foam.

¹H NMR: (DMSO-d₆) δ 0.94 (s, 9H), 1.43-1.74 (m, 8H), 2.18-2.23 (m, 1H),2.46-2.49 (m, 1H), 3.64 (s, 3H), 3.87 (d, J=12 Hz, 1H), 4.08-4.13 (m,1H), 4.42 (dd, J=7.5, 9.5 Hz, 1H), 4.78 (m, 1H), 5.30 (m, 1H), 7.03-7.05(m, 1H), 7.24 (s, 1H), 8.120 (d, J=6 Hz, 1H).

LC/MS rt-min (MH⁺): 2.34 (526, 528) (method C).

Step 5:

To a solution of the product of Step 4 (206 mg, 0.391 mmol) in THF (2mL) and methanol (2 mL) was added LiOH (28 mg, 1.2 mmol) in water (2mL). The mixture was stirred for 2 h at room temperature and quenchedwith 1N HCl until neutral pH. The organic volatiles were removed invacuo and to the residue was added buffer pH 4. The product wasextracted into EtOAc (2×20 mL) and dried (MgSO₄) to yield the titlecompound (196 mg, 100%) as a white solid.

¹H NMR: (methanol-d₄) δ 1.05 (s, 9H), 1.60-1.85 (m, 8H), 2.30-2.36 (m,1H), 2.62-2.67 (m, 1H), 3.97 (d, J=12 Hz, 1H), 4.26 (s, 1H), 4.32 (d,J=12 Hz, 1H), 4.58 (dd, J=7.5, 9.5 Hz, 1H), 4.89 (m (hidden), 1H), 5.29(m, 1H), 7.03-7.05 (m, 1H), 7.26 (s, 1H), 8.19 (d, J=6 Hz, 1H);

LC/MS rt-min (MH⁺): 2.17 (512, 514) (method B).

Step 6:

This product was prepared according to Example 420, Step 4 (reactiontime 20 h) in 20% yield starting from the product of Example 421, Step5.

¹H NMR: (methanol-d₄) δ 1.03 (s, 9H), 1.45-1.74 (m, 8H), 2.31-2.37 (m,1H), 2.65-2.69 (m, 1H), 3.99 (dd, J=12, 3.0 Hz, 1H), 4.26 (d, J=9.5 Hz,1H), 4.33 (d, J=11.5 Hz, 1H), 4.60 (dd, J=8.0, 9.5 Hz, 1H), 4.80 (m,1H), 5.35 (m, 1H), 6.72 (d, J=9.0 Hz, NH), 7.02 (dd, J=2.5, 6.0 Hz, 1H),7.39 (s, 1H), 7.50 (m, 3H (hidden)), 7.91 (d, J=6.5 Hz, 1H), 8.46 (d,J=6.0 Hz, 1H).

LC/MS rt-min (MH⁺): 2.03 (510) (method A).

Step 7:

Compound 421 was prepared according to Example 420, Step 6 in 68% yield,starting from the product of Example 421, Step 6.

¹H NMR: (methanol-d₄) δ 1.03 (s, 9H), 1.06-1.09 (m, 2H), 1.23-1.26 (m,2H), 1.43 (dd, J=5.5, 9.5 Hz, 1H), 1.47-1.77 (m, 8H), 1.88 (dd, J=5.5,8.5 Hz, 1H), 2.21-2.29 (m, 2H), 2.50-2.54 (m, 1H), 2.91-2.96 (m, 1H),4.06 (dd, J=3.0, 11.5 Hz, 1H), 4.28-4.30 (m, 2H), 4.44 (dd, J=7.0, 10.5Hz, 1H), 4.82 (m, 1H (hidden)), 5.12 (d, J=10 Hz, 1H), 5.30 (d, J=17 Hz,1H), 5.37 (m, 1H), 5.73-5.80 (m, 1H), 6.92 (d, J=9.5 Hz, NH), 6.98 (dd,J=5.5, 2.0 Hz, 1H), 7.37 (s, 1H), 7.43-7.50 (m, 3H), 7.90 (d, J=7.0 Hz,1H), 8.45 (d, J=5.5 Hz, 1H).

LC/MS rt-min (MH⁺): 2.37 (723) (method A).

Example 422 Preparation of Compound 422

Step 1:

To a mixture of the product of Example 420, Step 3 (102 mg, 0.198 mmol)and Pd(PPh₃)₄ (23 mg, 0.0198 mmol) in toluene (2 mL) was added4-tributylstannanyl-pyridine (87 mg, 0.24 mmol). The solution was heatedat 105° C. for 20 h under nitrogen. After cooling to room temperaturethe mixture was quenched with satd. NaHCO₃ and extracted with EtOAc(2×10 mL), and dried (Na₂SO₄). Purification using a Biotage 12 M column(eluted gradient hexane-acetone 20-60%) afforded the title compound (49mg, 49%) as a white solid.

¹H NMR: (methanol-d₄) δ 1.05 (s, 9H), 1.31 (s, 9H), 2.29-2.35 (m, 1H),2.68-2.72 (m, 1H), 3.77 (s, 3H), 4.00 (d, J=12 Hz, 1H), 4.24 (d, J=9.5Hz, 1H), 4.41 (d, J=12 Hz, 1H), 4.67 (dd, J=7.5, 10 Hz, 1H), 5.36 (m,1H), 6.51 (d, J=9.5 Hz, NH), 7.79-7.85 (m, 3H), 8.39 (s, 1H), 8.59 (s,1H), 8.67-8.68 (m, 2H).

LC/MS rt-min (MH⁺): 1.77 (514) (method C).

Step 2:

This product was prepared by the same procedure as described in Example420, Step in quantitative yield, except using the product from Example422, Step 1 instead.

¹H NMR: (methanol-d₄) δ 1.05 (s, 9H), 1.32 (s, 9H), 2.32-2.38 (m, 1H),2.69-2.74 (m, 1H), 3.99 (dd, J=3, 12 Hz, 1H), 4.25 (s, 1H), 4.40 (d,J=12 Hz, 1H), 4.64 (dd, J=7.5, 9.5 Hz, 1H), 5.36 (m, 1H), 7.85-7.87 (m,3H), 8.40 (s, 1H), 8.60 (s, 1H), 8.69-8.70 (m, 2H).

LC/MS rt-min (MH⁺): 1.57 (500) (method B).

Step 3:

Compound 422 was prepared by the same procedure as described in Example420, Step 6 in 51% yield as a white solid, except using the product fromExample 422, Step 2 instead.

¹H NMR: (DMSO-d₆) δ 0.95 (s, 9H), 1.03 (m, 1H), 1.08 (m, 1H), 1.23 (s,9H), 1.34-1.37 (m, 1H), 1.68-1.70 (m, 1H), 2.12-2.18 (m, 2H), 2.41-2.45(m, 1H), 2.93 (m, 1H), 3.90 (d, J=1 Hz, 1H), 4.06 (d, J=9 Hz, 1H), 4.17(d, J=1 Hz, 1H), 4.36 (dd, J=7, 10 Hz, 1H), 5.10 (d, J=12 Hz, 1H), 5.23(d, J=16.5 Hz, 1H), 5.40 (m, 1H), 5.59-5.66 (m, 1H), 6.60 (d, J=9 Hz,NH), 7.81-7.82 (m, 3H), 8.38 (s, 1H), 8.65 (s, 1H), 8.68-8.70 (m, 2H),8.93 (s, NH), 10.4 (s, NH).

LC/MS rt-min (MH⁺): 2.14 (711) (method D).

Example 423 Preparation of Compound 423

Step 1:

To a solution of the product of Example 420, Step 3 (1.00 g, 1.94 mmol)in THF (5 mL) and methanol (5 mL) was added LiOH (140 mg, 5.83 mmol) inwater (5 mL). The mixture was stirred for 2 h at room temperature andquenched with 1N HCl until neutral pH. The organic volatiles wereremoved in vacuo, buffer pH 4 was added and the product was extractedinto EtOAc (3×25 mL) and dried (MgSO₄) to yield the title compound (1.0g, 100%) as a white solid.

¹H NMR: (DMSO-d₆) δ 0.95 (s, 9H), 1.27 (s, 9H), 2.14-2.19 (m, 1H),2.47-2.50 (m, 1H), 3.80 (d, J=11.5 Hz, 1H), 3.99-4.07 (m, 2H), 4.15 (d,J=11.5 Hz, 1H), 4.36 (dd, J=8, 10 Hz, 1H), 5.25 (m, 1H), 6.66 (d, J=9Hz, NH), 7.75 (s, 1H), 8.28 (s, 1H), 8.31 (s, 1H), 12.5 (s, 1H).

LC/MS rt-min (MH⁺): 2.47 (500, 502) (method A).

Step 2:

Compound 423 was accomplished according to Example 420, Step 6 in 52%yield, starting from the products of Example 423, Step 1 and Example 8,Step 3 (racemic Pi, (1R,2S) and (1S,2R)).

¹H NMR: (DMSO-d₆) δ 0.93, 0.95 (s, 9H), 1.00-1.09 (m, 4H), 1.28, 1.29(s, 9H), 1.37-1.39 (m, 1H), 1.69-1.72 (m, 1H), 2.09-2.22 (m, 2H),2.36-2.46 (m, 1H), 2.88-2.94 (m, 1H), 3.82-3.87 (m, 1H), 4.02-4.04 (m,1H), 4.10-4.15 (m, 1H), 4.31-4.34 (m, 1H), 5.10 (d, J=10.5 Hz, 1H),5.22-5.28 (m, 1H), 5.54-5.66 (m, 1H), 6.56, 6.60 (d, J=9.5 Hz, NH),7.74, 7.76 (s, 1H), 8.28-8.29 (m, 1H), 8.32 (s, 1H), 8.79, 8.89 (s, 1H).

LC/MS rt-min (MH⁺): 2.42 (712, 714) (method B).

Example 424 Preparation of Compound 424

Step 1:

To a solution of the product of Example 423, Step 1 (91 mg, 0.18 mmol),Pd(PPh₃)₄ (10.5 mg, 0.0091 mmol), and 3-furyl boronic acid (25.4 mg,0.227 mmol) in DMF (2 mL) was added 2M aqueous Na₂CO₃ (0.273 mL, 0.546mmol). The mixture was heated at 110° C. for 2.5 h under nitrogen. Aftercooling to room temperature the solid was removed by filtration, and thefiltrate concentrated in vacuo. The residue was purified by preparativeHPLC (gradient 30-80% B) to afford the title compound (64 mg, 72%) as awhite solid.

¹H NMR: (DMSO-d₆) δ 0.95 (s, 9H), 1.25 (s, 9H), 2.17-2.21 (m, 1H), 2.50(m (hidden), 1H), 3.84-3.86 (m, 1H), 4.09-4.15 (m, 2H), 4.37 (t, J=9 Hz,1H), 5.28 (m, 1H), 6.67 (d, J=9 Hz, NH), 7.08 (s, 1H), 7.62 (s, 1H),7.79 (s, 1H), 8.17 (s, 1H), 8.33 (s, 1H), 8.51 (s, 1H), 12.6 (s, 1H).

LC/MS rt-min (MH⁺): 1.60 (488) (method B).

Step 2:

Compound 424 was prepared according to Example 420, Step 6 in 55% yield,starting from the product of Example 424, Step 1.

¹H NMR: (methanol-d₄) δ 1.02 (s, 9H), 1.05-1.09 (m, 2H), 1.22-1.25 (m,2H), 1.34 (s, 9H), 1.42-1.45 (m, 1H), 1.86-1.89 (m, 1H), 2.21-2.26 (m,1H), 2.48-2.52 (m, 1H), 2.91-2.96 (m, 1H), 4.02 (d, J=12 Hz, 1H),4.24-4.29 (m, 2H), 4.43-4.47 (dd, J=7.5, 10.5 Hz, 1H), 5.12 (d, J=10.5Hz, 1H), 5.28-5.32 (m, 1H), 5.73-5.81 (m, 1H), 6.63 (d, J=9 Hz, NH),6.90 (s, 1H), 7.60-7.62 (m, 2H), 8.07 (s, 1H), 8.12 (s, 1H), 8.39 (s,NH).

LC/MS rt-min (MH⁺): 2.30 (700) (method E).

Example 425

Preparation of Compound 425

Compound 425 was accomplished according to Example 421, Step 7 in 45%yield, starting from the product of Example 421, Step 5.

¹H NMR: (DMSO-d₆) δ 0.95 (s, 9H), 1.03-1.04 (m, 1H), 1.08-1.09 (m, 1H),1.33-1.36 (m, 1H), 1.46-1.75 (m, 9H), 2.08-2.12 (m, 1H), 2.17 (q, J=9Hz, 1H), 2.35-2.38 (m, 1H), 2.88-2.93 (m, 1H), 3.91 (d, J=9.5 Hz, 1H),4.05-4.10 (m, 2H), 4.27 (dd, J=10, 7 Hz, 1H), 4.80 (m, 1H), 5.10 (d,J=12 Hz, 1H), 5.23 (d, J=16.5 Hz, 1H), 5.32 (m, 1H), 5.58-5.66 (m, 1H),6.94 (d, J=9 Hz, NH), 7.03-7.05 (m, 1H), 7.26 (s, 1H), 8.21 (d, J=6 Hz,1H), 8.86 (s, NH), 10.4 (s, NH).

LC/MS rt-min (MH⁺): 2.56 (724, 726) (method D).

Example 426 Preparation of Compound 426

Step 1:

This product was prepared according to Example 421, Step 6 in 65% yield,except using 3-furanoboronic acid.

¹H NMR: (methanol-d₄) δ 1.03 (s, 9H), 1.49-1.76 (m, 8H), 2.30-2.35 (m,1H), 2.63-2.67 (m, 1H), 3.98 (d, J=12 Hz, 1H), 4.26 (t, J=9 Hz, 1H),4.31 (d, J=12 Hz, 1H), 4.58 (t, J=8.0 Hz, 1H), 5.32 (m, 1H), 6.71 (d,J=9.5 Hz, NH), 6.92 (dd, J=2.5, 6.0 Hz, 1H), 6.98 (s, 1H), 7.25 (d,J=2.5 Hz, 1H), 7.60 (s, 1H), 8.15 (s, 1H), 8.35 (d, J=5.5 Hz, 1H).

LC/MS rt-min (MH⁺): 1.94 (500) (method D).

Step 2:

Compound 426 was prepared according to Example 421, Step 7 in 57% yield,starting from the product of Example 426, Step 1.

¹H NMR: (DMSO-d₆) δ 0.95 (s, 9H), 1.03-1.04 (m, 2H), 1.08-1.09 (m, 2H),1.34-1.71 (m, 8H), 2.09-2.20 (m, 2H), 2.37-2.41 (m, 1H), 2.93 (m, 1H),3.96 (d, J=9.5 Hz, 1H), 4.07-4.11 (m, 2H), 4.29 (m, 1H), 4.78 (m, 1H),5.10 (d, J=10.5 Hz, 1H), 5.23 (d, J=17 Hz, 1H), 5.34 (m, 1H), 5.59-5.66(m, 1H), 6.87-6.91 (m, 2H), 7.08 (s, 1H), 7.27 (s, 1H), 7.75 (s, 1H),8.33 (s, 1H), 8.39 (d, J=6 Hz, 1H), 8.90 (s, NH), 10.4 (s, 1H).

LC/MS rt-min (MH⁺): 2.38 (712) (method D).

Example 427 Preparation of Compound 427

Step 1:

This product was prepared according to Example 421, Step 6 in 86% yield,except using 4-fluorophenylboronic acid.

¹H NMR: (methanol-d₄) δ 1.03 (s, 9H), 1.44-1.75 (m, 8H), 2.30-2.36 (m,1H), 2.64-2.69 (m, 1H), 3.99 (dd, J=12, 3.5 Hz, 1H), 4.26 (d, J=9.5 Hz,1H), 4.32 (d, J=12 Hz, 1H), 4.59 (dd, J=8.0, 9.5 Hz, 1H), 4.81 (m, 1H),5.34 (m, 1H), 6.73 (d, J=9.5 Hz, NH), 6.99 (dd, J=2.5, 6.0 Hz, 1H), 7.20(t, 2H (hidden)), 7.36 (s, 1H (hidden)), 7.94-7.97 (m, 2H), 8.44 (d,J=6.0 Hz, 1H).

LC/MS rt-min (MH⁺): 2.22 (528) (method A).

Step 2:

Compound 427 was prepared according to Example 421, Step 7 in 53% yield,starting from the product of Example 427, Step 1.

¹H NMR: (methanol-d₄) δ 1.03 (s, 9H), 1.06-1.09 (m, 2H), 1.23-1.26 (m,2H), 1.43 (dd, J=5.0, 9.5 Hz, 1H), 1.47-1.78 (m, 8H), 1.88 (dd, J=5.5,8.0 Hz, 1H), 2.21-2.29 (m, 2H), 2.49-2.53 (m, 1H), 2.91-2.96 (m, 1H),4.05 (dd, J=3.0, 11.5 Hz, 1H), 4.27-4.29 (m, 2H), 4.44 (dd, J=7.0, 10.5Hz, 1H), 4.83 (m, 1H (hidden)), 5.12 (d, J=10 Hz, 1H), 5.29 (d, J=17 Hz,1H), 5.37 (m, 1H), 5.73-5.80 (m, 1H), 6.93 (d, J=9.5 Hz, NH), 6.98 (m,1H), 7.21 (t, J=9.0 Hz, 2H), 7.36 (s, 1H), 7.94-7.97 (m, 2H), 8.44 (d,J=6.0 Hz, 1H).

LC/MS rt-min (MH⁺): 2.42 (741) (method A).

Example 428 Preparation of Compound 428

Step 1:

This product was prepared according to Example 421, Step 6 in 37% yield,except using 4-methoxyphenylboronic acid.

¹H NMR: (methanol-d₄) δ 1.03 (s, 9H), 1.45-1.74 (m, 8H), 2.31-2.36 (m,1H), 2.64-2.68 (m, 1H), 3.85 (s, 3H), 3.98 (dd, J=12, 3.5 Hz, 1H), 4.25(d, J=9.0 Hz, 1H), 4.33 (d, J=11.5 Hz, 1H), 4.59 (dd, J=8.0, 10 Hz, 1H),4.77 (m, 1H), 5.36 (m, 1H), 6.72 (d, J=9.0 Hz, NH), 7.01 (m, 1H), 7.04(d, J=9.0 Hz, 2H), 7.37 (d, J=2.0 Hz), 7.86 (d, J=9.0 Hz, 2H), 8.42 (d,J=6.0 Hz, 1H).

LC/MS rt-min (MH⁺): 2.09 (540) (method A).

Step 2:

Compound 428 was prepared according to Example 420, Step 6 in 72% yield,starting from the product of Example 428, Step 1.

¹H NMR: (methanol-d₄) δ 1.03 (s, 9H), 1.06-1.09 (m, 2H), 1.23-1.25 (m,2H), 1.43 (dd, J=5.5, 9.5 Hz, 1H), 1.48-1.77 (m, 8H), 1.88 (dd, J=5.0,8.0 Hz, 1H), 2.21-2.28 (m, 2H), 2.50-2.54 (m, 1H), 2.91-2.96 (m, 1H),3.85 (s, 3H), 4.05 (d, J=12 Hz, 1H), 4.27-4.29 (m, 2H), 4.44 (dd, J=7.0,10.5 Hz, 1H), 4.84 (m, 1H (hidden)), 5.12 (d, J=10 Hz, 1H), 5.30 (d,J=17 Hz, 1H), 5.37 (m, 1H), 5.72-5.80 (m, 1H), 6.89 (d, J=9.5 Hz, NH),6.95 (dd, J=2.5, 6.0 Hz, 1H), 7.03 (d, J=9.0 Hz, 1H), 7.33 (d, J=2.5 Hz,1H), 7.86 (d, J=9.0 Hz, 2H), 8.41 (d, J=6.0 Hz, 1H).

LC/MS rt-min (MH⁺): 2.40 (753) (method A).

Example 429 Preparation of Compound 429

Step 1:

This product was prepared according to Example 421, Step 6 in 22% yield,except using 2-thiopheneboronic acid.

¹H NMR: (methanol-d₄) δ 1.03 (s, 9H), 1.46-1.74 (m, 8H), 2.29-2.35 (m,1H), 2.63-2.67 (m, 1H), 3.97 (d, J=12 Hz, 1H), 4.26 (d, J=8.5 Hz, 1H),4.31 (d, J=12 Hz, 1H), 4.60 (t, J=8.5 Hz, 1H), 4.81 (m, 1H (hidden)),5.31 (m, 1H), 6.87-6.89 (m, 2H), 7.13 (d, J=5.0 Hz, 1H), 7.52 (s, 1H(hidden)), 7.70 (d, J=2.5 Hz, 1H), 8.32 (d, J=6.0 Hz, 1H).

LC/MS rt-min (MH⁺): 1.96 (516) (method A).

Step 2:

Compound 429 was prepared according to Example 420, Step 6 in 57% yield,starting from the product of Example 429, Step 1.

¹H NMR: (methanol-d₄) δ 1.02 (s, 9H), 1.07 (m, 2H), 1.24 (m, 2H), 1.43(dd, J=5.5, 9.5 Hz, 1H), 1.53-1.78 (m, 8H), 1.88 (dd, J=5.5, 8.0 Hz,1H), 2.21-2.28 (m, 2H), 2.49-2.53 (m, 1H), 2.92-2.96 (m, 1H), 4.04 (d,J=10 Hz, 1H), 4.26-4.29 (m, 2H), 4.43 (t, J=9.0 Hz, 1H), 4.82 (m, 1H(hidden)), 5.12 (d, J=10.5 Hz, 1H), 5.29 (d, J=17.5 Hz, 1H), 5.35 (m,1H), 5.73-5.80 (m, 1H), 6.89 (d, J=4.5 Hz, 1H), 6.92 (d, J=4.5 Hz, NH),7.13 (s, 1H), 7.35 (s, 1H), 7.51 (d, J=4.5 Hz, 1H), 7.70 (s, 1H), 8.32(d, J=5.5 Hz, 1H).

LC/MS rt-min (MH⁺): 2.37 (728) (method A).

Example 430 Preparation of Compound 430

Step 1:

This product was prepared according to Example 421, Step 6 in 17% yield,except using 3-thiopheneboronic acid.

¹H NMR: (methanol-d₄) δ 1.03 (s, 9H), 1.43-1.74 (m, 8H), 2.30-2.36 (m,1H), 2.64-2.68 (m, 1H), 3.98 (dd, J=11.5, 3.0 Hz, 1H), 4.25 (d, J=9.0Hz, 1H), 4.32 (d, J=11.5 Hz, 1H), 4.60 (dd, J=8.0, 9.5 Hz, 1H), 4.80 (m,1H (hidden)), 5.33 (m, 1H), 6.96 (dd, J=2.5, 6.0 Hz, 1H), 7.36 (s, 1H(hidden)), 7.51 (s, 1H (hidden)), 7.67 (d, J=5.0 Hz, 1H), 8.03 (s, 1H),8.39 (d, J=6.0 Hz, 1H).

LC/MS rt-min (MH⁺): 1.94 (516) (method A).

Step 2:

Compound 430 was prepared according to Example 420, Step 6 in 45% yield,starting from the product of Example 430, Step 1.

¹H NMR: (methanol-d₄) δ 1.02 (s, 9H), 1.06 (m, 2H), 1.29 (m, 2H), 1.43(dd, J=5.5, 9.5 Hz, 1H), 1.43-1.75 (m, 8H), 1.87 (dd, J=6.0, 7.5 Hz,1H), 2.20-2.27 (m, 2H), 2.49-2.53 (m, 1H), 2.93-2.95 (m, 1H), 4.05 (d,J=9.0 Hz, 1H), 4.27-4.29 (m, 2H), 4.42-4.45 (m, 1H), 4.85 (m, 1H(hidden)), 5.12 (d, J=10.0 Hz, 1H), 5.29 (d, J=17.5 Hz, 1H), 5.36 (m,1H), 5.73-5.80 (m, 1H), 6.92-6.94 (m, 2H), 7.35 (s, 1H), 7.51 (s, 1H),7.66 (d, J=4.5 Hz, 1H), 8.00 (s, 1H), 8.38 (d, J=5.5 Hz, 1H).

LC/MS rt-min (MH⁺): 2.36 (728) (method A).

Example 431 Preparation of Compound 431

Step 1:

To a mixture of the product of Example 421, Step 5 (100 mg, 0.195 mmol)and Pd(PPh₃)₄ (23 mg, 0.0195 mmol) in dioxane (3 mL) was added2-tributylstannylthiazole (95 mg, 0.254 mmol) and triethylamine (82 μL,0.585 mmol). The solution was heated at 95° C. for 5 h under nitrogen,then at 105° C. for 15 h. After cooling to room temperature the mixturewas filtered and concentrated. The residue was purified by preparativeHPLC (gradient 30-80% B). The combined fractions were partitionedbetween buffer pH 4 and dichloromethane. The aqueous phase was extractedwith dichloromethane and the combined organic extracts washed with brineand dried (MgSO₄). The title compound (31 mg) was obtained as acolorless oil, significantly contaminated with tributylstannyl residue.

¹H NMR: (methanol-d₄) δ 1.03 (s, 9H), 1.4 (m, 8H (hidden)), 2.32-2.36(m, 1H), 2.64-2.68 (m, 1H), 4.00 (d, J=11.5 Hz, 1H), 4.25 (s, 1H), 4.33(d, J=11.5 Hz, 1H), 4.60 (t, J=8.5 Hz, 1H), 4.80 (m, 1H (hidden)), 5.33(m, 1H), 7.03 (br s, 1H), 7.70 (s, 1H), 7.73 (s, 1H), 7.93 (s, 1H), 8.41(d, J=5.5 Hz, 1H).

LC/MS rt-min (MH⁺): 2.25 (517) (method A).

Step 2:

Compound 431 was prepared according to Example 420, Step 6 in 41% yield,starting from the product of Example 431, Step 1.

¹H NMR: (methanol-d₄) δ 1.02 (s, 9H), 1.06-1.70 (m, 13H), 1.86-1.89 (m,1H), 2.22-2.31 (m, 2H), 2.51-2.55 (m, 1H), 2.92-2.94 (m, 1H), 4.06 (d,J=11.5 Hz, 1H), 4.23-4.32 (m, 2H), 4.44-4.47 (m, 1H), 4.82 (m, 1H(hidden)), 5.12 (d, J=11 Hz, 1H), 5.30 (d, J=17 Hz, 1H), 5.36 (m, 1H),5.73-5.81 (m, 1H), 6.90 (d, J=9.0 Hz, NH), 7.04 (m, 1H), 7.71 (m, 2H),7.93 (s, 1H), 8.42 (d, J=5.5 Hz, 1H).

LC/MS rt-min (MH⁺): 2.46 (729) (method A).

Example 432 Preparation of Compound 432

Step 1:

To a mixture of the product of Example 423, Step 1 (102 mg, 0.204 mmol)and Pd(PPh₃)₄ (24 mg, 0.0204 mmol) in dioxane (3 mL) was added2-tributylstannylthiazole (99 mg, 0.265 mmol) and triethylamine (85 μL,0.612 mmol). The solution was heated at 95° C. for 5 h under nitrogen,then at 105° C. for 15 h. After cooling to room temperature the mixturewas filtered and concentrated. The residue was purified by preparativeHPLC (gradient 30-80% B). The combined fractions were neutralized withconc. ammonia and concentrated. The residue was partitioned betweenbuffer pH 4 and dichloromethane. The aqueous phase was extracted withdichloromethane and the combined organic extracts washed with brine anddried (MgSO₄). The title compound (33 mg) was obtained as a colorlessoil, significantly contaminated with tributylstannyl containing residue.

¹H NMR: (methanol-d₄) δ 1.03 (s, 9H), 1.30 (s, 9H), 2.28-2.32 (m, 1H),2.62-2.67 (m, 1H), 3.98 (d, J=11.5 Hz, 1H), 4.21 (s, 1H), 4.34 (d,J=11.5 Hz, 1H), 4.58 (t, J=9.0 Hz, 1H), 5.31 (m, 1H), 7.71 (d, J=2.5 Hz,1H), 7.94-7.96 (m, 2H), 8.33 (s, 1H), 8.73 (s, 1H).

LC/MS rt-min (MH-Boc): 2.21 (405) (method A).

Step 2:

Compound 432 was prepared according to Example 420, Step 6 in 42% yield,starting from the product of Example 432, Step 1.

¹H NMR: (methanol-d₄) δ 1.02 (s, 9H), 1.07-1.08 (m, 2H), 1.24 (m, 2H),1.32 (s, 9H), 1.44 (m, 1H (hidden)), 1.86-1.89 (m, 1H), 2.21-2.29 (m,2H), 2.51-2.55 (m, 1H), 2.93-2.95 (m, 1H), 4.04 (d, J=12 Hz, 1H), 4.23(d, J=9.5 Hz, 1H), 4.33 (d, J=12 Hz, 1H), 4.47 (t, J=9.5 Hz, 1H), 5.12(d, J=10.0 Hz, 1H), 5.30 (d, J=18 Hz, 1H), 5.36 (m, 1H), 5.72-5.81 (m,1H), 6.62 (d, J=8.5 Hz, NH), 7.73 (s, 1H), 7.96 (m, 1H), 8.34 (s, 1H),8.74 (s, 1H).

LC/MS rt-min (MH⁺): 2.42 (717) (method A).

Example 433 Preparation of Compound 433

Step 1:

To a mixture of the product of Example 420, Step 3 (1.00 g, 1.94 mmol)and Pd(PPh₃)₄ (112 mg, 0.097 mmol) in dioxane (15 mL) was addedtributyl(1-ethoxyvinyl)tin (876 mg, 2.43 mmol. The solution was heatedat 105° C. for 6 h under nitrogen. After cooling to room temperature themixture was filtered and concentrated. The residue was partitionedbetween satd NaHCO₃ and ethyl acetate. The aqueous phase was extractedwith ethyl acetate and the combined organic extracts washed with 5% aq.KF and brine and dried (MgSO₄). Purification using a Biotage 40 M column(eluted gradient hexane-EtOAc 40-70%) afforded the title compound (624mg, 64%) as a yellow oil.

¹H NMR: (DMSO-d₆) δ 0.95 (s, 9H), 1.27 (s, 9H), 1.35 (t, J=7.0 Hz, 3H),2.16-2.21 (m, 1H), 2.50 (m, 1H (hidden)), 3.64 (s, 3H), 3.85 (d, J=11Hz, 1H), 3.90 (q, J=7.0 Hz, 2H), 4.09 (d, J=9.0 Hz, 1H), 4.13 (d, J=11Hz, 1H), 4.40 (d, J=2.5 Hz, 1H), 4.45 (t, J=8.0 Hz, 1H), 4.91 (d, J=2.5Hz, 1H), 5.27 (m, 1H), 6.69 (d, J=9.0 Hz, NH), 7.48 (s, 1H), 8.25 (s,1H), 8.46 (s, 1H).

LC/MS rt-min (MH⁺): 2.14 (507) (method B).

Step 2:

To a solution of the product of Example 433, Step 1 (125 mg, 0.247) inTHF (3 mL) and water (111 μL, 6.18 mmol) was added NBS (44 mg, 0.247mmol). After stirring at room temperature for 20 min. the mixture wasconcentrated and partitioned between ethyl acetate and brine. Theorganic phase was dried (MgSO₄) to give the intermediate bromomethylketone. This intermediate was dissolved in DMF and treated withthioacetamide (24 mg, 0.321 mmol) and NaHCO₃ (31 mg, 0.371 mmol). Themixture was stirred for 2 h at room temperature, concentrated andsuspended in satd. NaHCO₃. The product was extracted with ethyl acetate(2×), washed with brine, and dried (MgSO₄). Purification using a Biotage12 M column (eluted gradient hexane-EtOAc 50-70%) afforded the titlecompound (50 mg, 38%) as a pale oil.

¹H NMR: (methanol-d₄) δ 1.02 (s, 9H), 1.31 (s, 9H), 2.27-2.31 (m, 1H),2.63-2.67 (m, 1H), 2.77 (s, 3H), 3.74 (s, 3H), 3.98 (d, J=10.0 Hz, 1H),4.22 (d, J=9.5 Hz, 1H), 4.34 (d, J=10.5 Hz, 1H), 4.64 (t, J=9.0 Hz, 1H),5.29 (m, 1H), 6.38 (s, NH), 7.87 (s, 1H), 7.91 (s, 1H), 8.20 (s, 1H),8.71 (s, 1H).

LC/MS rt-min (MH⁺): 1.96 (533) (method B).

Step 3:

This product was prepared according to Example 420, Step 5, except usingthe product of Example 433, Step 2 instead.

¹H NMR: (DMSO-d₆) δ 0.96 (s, 9H), 1.25 (s, 9H), 2.18-2.23 (m, 1H), 2.50(m, 1H (hidden)), 2.73 (s, 3H), 3.86 (d, J=11.5 Hz, 1H), 4.10 (d, J=9.0Hz, 1H), 4.14 (d, J=11.5 Hz, 1H), 4.39 (t, J=8.5 Hz, 1H), 5.30 (m, 1H),6.60 (br s, NH), 7.84 (s, 1H), 8.14 (s, 1H), 8.24 (s, 1H), 8.78 (s, 1H).

LC/MS rt-min (MH⁺): 1.84 (519) (method B).

Step 4:

Compound 433 was prepared according to Example 420, Step 6 in 46% yield,starting from the product of Example 433, Step 3.

¹H NMR: (methanol-d₄) δ 1.05 (s, 9H), 1.09-1.11 (m, 2H), 1.26-1.29 (m,2H), 1.36 (s, 9H), 1.45-1.48 (m, 1H), 1.89-1.92 (m, 1H), 2.25-2.30 (m,2H), 2.53-2.57 (m, 1H), 2.80 (s, 3H), 2.94-3.00 (m, 1H), 4.06 (d, J=10.5Hz, 1H), 4.27 (s, 1H), 4.32 (d, J=12 Hz, 1H), 4.48 (dd, J=10.5, 7.0 Hz,1H), 5.15 (d, J=10.5 Hz, 1H), 5.33 (d, J=17 Hz, 1H), 5.35 (m, 1H),5.76-5.83 (m, 1H), 7.93 (s, 1H), 7.95 (m, 1H), 8.22 (s, 1H), 8.74 (s,1H).

LC/MS rt-min (MH⁺): 2.21 (732) (method B).

Example 434 Preparation of Compound 434

Step 1:

To a solution of the product of Example 421, Step 1 (300 mg, 1.56 mmol)and Pd(PPh₃)₄ (90 mg, 0.078 mmol) in DMF (6 mL) was added1-methyl-2-(tributylstannyl)-1H-pyrrole (750 mg, 2.03 mmol) andtriethylamine (0.435 mL, 3.12 mmol). The solution was heated at 150° C.for 30 min under nitrogen in a microwave oven (Emrys, PersonalChemistry). After cooling to room temperature the mixture was dilutedwith diethyl ether and 5% aq. KF and filtered. The aqueous phase wasextracted with diethyl ether (2×). The combined organic extracts werewashed with 5% aq. KF, water and brine and dried (MgSO₄). Purificationusing a Biotage 25 S column (eluted gradient hexane-diethyl ether 0-5%)afforded the title compound (169 mg, 56%) as a colorless oil.

¹H NMR: (DMSO-d₆) δ 3.94 (s, 3H), 6.10 (s, 1H), 6.77 (d, J=2.0 Hz, 1H),6.93 (s, 1H), 7.27 (d, J=4.0 Hz, 1H), 7.77 (s, 1H), 8.49 (d, J=5.0 Hz,1H),

LC/MS rt-min (MH⁺): 1.15 (193, 195) (method B).

Step 2:

This product was prepared according to Example 421, Step 2 in 39% yield,starting from the product of Example 434, Step 1.

¹H NMR: (methanol-d₄) δ 1.42, 1.43 (s, 9H (rotamers)), 2.27-2.34 (m,1H), 2.55-2.62 (m, 1H), 3.75 (m, 2H), 3.76, 3.74 (s, 3H (rotamers)),3.84 (s, 3H), 4.39-4.45 (m, 1H), 5.19 (m, 1H), 6.10 (m, 1H), 6.49 (m,1H), 6.78 (s, 1H), 6.80-6.82 (m, 1H), 7.07 (s, 1H), 8.35 (d, J=6.0 Hz,1H).

LC/MS rt-min (MH⁺, carboxylic acid): 1.49 (389) (method B).

Step 3:

The product of Example 434, Step 2 (90 mg, 0.22 mmol) was dissolved indichloromethane (1.5 mL) and TFA (1.0 mL, 9.0 mmol). The solution wasstirred at room temperature for 45 min., and concentrated. The residuewas treated with 1N HCl in diethyl ether (5 mL) and concentrated. Thetitle compound was obtained in quantitative yield as a pale oil.

LC/MS rt-min (MH⁺): 0.32 (302) (method B).

Step 4:

This product was prepared according to Example 420, Step 3 in 80% yield,starting from the product of Example 434, Step 3.

¹H NMR: (methanol-d₄) δ 1.02 (s, 9H), 1.31 (s, 9H), 2.25-2.31 (m, 1H),2.61-2.64 (m, 1H), 3.73 (s, 3H), 3.84 (s, 3H), 3.97-3.99 (m, 1H), 4.22(d, J=9.5 Hz, 1H), 4.32 (d, J=11.5 Hz, 1H), 4.61 (t, J=7.5 Hz, 1H), 5.27(m, 1H), 6.10 (m, 1H), 6.42 (br d, J=9.0 Hz, NH), 6.49 (m, 1H), 6.78 (m,1H), 6.81 (m, 1H), 7.07 (s, 1H), 8.34 (d, J=5.5 Hz, 1H).

LC/MS rt-min (MH⁺): 1.81 (516) (method B).

Step 5:

To a solution of the product of Example 434, Step 4 (92 mg, 0.179 mmol)in THF (1 mL) and methanol (1 mL) was added LiOH (13 mg, 0.536 mmol) inwater (1 mL). The mixture was stirred for 1.5 h at room temperature andquenched with 1N HCl until neutral pH. The organic volatiles wereremoved in vacuo, and the residue purified by preparative HPLC (gradient10-80% B). The combined fractions were neutralized with conc. ammoniaand concentrated. The residue was partitioned between buffer pH 4 andethyl acetate. The aqueous phase was extracted with ethyl acetate andthe combined organic extracts washed with brine and dried (MgSO₄). Thetitle compound (56 mg, 62%) was obtained as a white solid.

¹H NMR: (methanol-d₄) δ 1.03 (s, 9H), 1.32 (s, 9H), 2.31-2.35 (m, 1H),2.62-2.67 (m, 1H), 3.85 (s, 3H), 3.98 (d, J=11.5 Hz, 1H), 4.21-4.23 (m,1H), 4.33 (d, J=11.5 Hz, 1H), 4.58 (t, J=8.0 Hz, 1H), 5.31 (m, 1H), 6.12(m, 1H), 6.40 (br d, J=8.0 Hz, NH), 6.52 (m, 1H), 6.82 (m, 1H), 6.87 (m,1H), 7.12 (s, 1H), 8.36 (d, J=5.5 Hz, 1H).

LC/MS rt-min (MH⁺): 1.73 (502) (method B).

Step 6:

Compound 434 was prepared according to Example 420, Step 6 in 68% yield,starting from the products of Example 434, Step 5.

¹H NMR: (methanol-d₄) δ 1.02 (s, 9H), 1.06-1.09 (m, 2H), 1.23-1.26 (m,2H), 1.33 (s, 9H), 1.42-1.45 (m, 1H), 1.86-1.89 (m, 1H), 2.21-2.27 (m,2H), 2.47-2.51 (m, 1H), 2.91-2.96 (m, 1H), 3.85 (s, 3H), 4.04 (d, J=12Hz, 1H), 4.24 (d, J=10.0 Hz, 1H), 4.28 (d, J=12 Hz, 1H), 4.43 (dd,J=10.0, 7.0 Hz, 1H), 5.12 (d, J=10.0 Hz, 1H), 5.30 (d, J=17 Hz, 1H),5.32 (m, 1H), 5.73-5.80 (m, 1H), 6.10 (m, 1H), 6.49 (m, 1H), 6.64 (br d,J=9.0 Hz, NH), 6.79 (m, 1H), 6.82 (m, 1H), 7.08 (s, 1H), 8.35 (d, J=5.5Hz, 1H).

LC/MS rt-min (MH⁺): 2.08 (714) (method B).

Example 435 Preparation of Compound 435

Step 1:

This product was prepared according to Example 421, Step 2 in 74% yield,starting from 2,6-dibromopyridine

¹H NMR: (DMSO-d₆) δ 1.34, 1.38 (s, 9H (rotamers)), 2.23-2.31 (m, 1H),2.43-2.47 (m, 1H (hidden)), 3.53 (d, J=12 Hz, 1H), 3.66, 3.69 (s, 3H(rotamers)), 3.72-3.75 (m, 1H), 4.29-4.34 (m, 1H), 5.42 (m, 1H), 6.89(d, J=7.5 Hz, 1H), 7.26 (d, J=7.5 Hz, 1H), 7.68 (t, J=7.5 Hz, 1H).

LC/MS rt-min (MH⁺): 2.62 (523, 425) (method A).

Step 2:

This product was prepared according to Example 420, Step 2 inquantitative yield, starting from the product of Example 435, Step 1.

LC/MS rt-min (MH⁺): 1.42 (301, 303) (method B).

Step 3:

This product was prepared according to Example 420, Step 3 in 96% yield,starting from the product of Example 435, Step 2.

¹H NMR: (DMSO-d₆) δ 0.95 (s, 9H), 1.28 (s, 9H), 2.20-2.26 (m, 1H),2.45-2.48 (m, 1H), 3.64 (s, 3H), 3.94 (d, J=9.5 Hz, 1H), 4.01-4.08 (m,2H), 4.45 (t, J=8.5 Hz, 1H), 5.53 (m, 1H), 6.66 (d, J=7.0 Hz, 1H), 6.82(d, J=7.0 Hz, 1H), 7.25 (d, J=7.0 Hz, 1H), 7.64-7.69 (m, 1H).

LC/MS rt-min (MH⁺): 2.73 (514, 516) (method A).

Step 3:

This product was prepared according to Example 420, Step 5 inquantitative yield, starting from the product of Example 435, Step 2.

¹H NMR: (DMSO-d₆) δ 0.95 (s, 9H), 1.28 (s, 9H), 2.19-2.25 (m, 1H),2.43-2.47 (m, 1H), 3.94 (m, 1H), 4.01-4.08 (m, 2H), 4.36 (t, J=8.5 Hz,1H), 5.52 (m, 1H), 6.65 (d, J=8.0 Hz, 1H), 6.82 (d, J=7.5 Hz, 1H), 7.25(d, J=7.5 Hz, 1H), 7.67 (t, J=7.5 Hz, 1H), 12.6 (s, 1H).

LC/MS rt-min (MNa⁺): 2.51 (522, 524) (method B).

Step 4:

To a solution of the product of Example 435, Step 3 (125 mg, 0.250mmol), Pd(PPh₃)₄ (14.4 mg, 0.0125 mmol), and 3-furyl boronic acid (35mg, 0.313 mmol) in DMF (2 mL) and water (0.025 mL) was added Cs₂CO₃ (244mg, 0.750 mmol). The mixture was heated at 105° C. for 3 h undernitrogen. After cooling to room temperature the solid was removed byfiltration, and the filtrate concentrated in vacuo. The residue waspurified by preparative HPLC (gradient 30-100% B). The combinedfractions were neutralized with conc. ammonia and concentrated. Theresidue was partitioned between buffer pH 4 and dichloromethane. Theaqueous phase was extracted with dichloromethane and the combinedorganic extracts washed with brine and dried (MgSO₄). The title compound(90 mg, 76%) was obtained as a white foam.

¹H NMR: (methanol-d₄) δ 1.06 (s, 9H), 1.39 (s, 9H), 2.36-2.42 (m, 1H),2.61-2.65 (m, 1H), 4.12 (dd, J=4.0.11 Hz, 1H), 4.19 (d, J=11 Hz, 1H),4.28 (s, 1H), 4.62 (t, J=8.5 Hz, 1H), 5.79 (m, 1H), 6.64 (d, J=8.0 Hz,1H), 6.96 (s, 1H), 7.22 (d, J=8.0 Hz, 1H), 7.58 (m, 1H), 7.66 (t, J=8.0Hz, 1H), 8.13 (s, 1H).

LC/MS rt-min (MNa⁺): 2.53 (511) (method B).

Step 5:

Compound 435 was prepared according to Example 420, Step 6 in 57% yield,starting from the product of Example 435, Step 4.

¹H NMR: (DMSO-d₆) δ 0.96 (s, 9H), 1.02-1.05 (m, 2H), 1.09-1.11 (m, 2H),1.19 (s, 9H), 1.35-1.38 (m, 1H), 1.71 (dd, J=5.5, 8.0 Hz, 1H), 2.15-2.22(m, 2H), 2.37-2.41 (m, 1H), 2.93 (br m, 1H), 4.03-4.08 (m, 3H), 4.35 (brt, 1H), 5.10 (d, J=10.5 Hz, 1H), 5.24 (d, J=17 Hz, 1H), 5.60-5.67 (m,1H), 5.73 (m, 1H), 6.47 (br s, 1H), 6.64 (d, J=7.5 Hz, 1H), 7.04 (s,1H), 7.31 (d, J=7.5 Hz, 1H), 7.72 (t, J=7.5 Hz, 1H), 7.77 (s, 1H), 8.32(s, 1H), 8.92 (s, NH), 10.4 (s, NH).

LC/MS rt-min (MNa⁺): 2.64 (723) (method B).

Example 436 Preparation of Compound 436

Step 1:

This product was prepared according to Example 435, Step 4 in 73% yield,except using phenylboronic acid instead.

¹H NMR: (methanol-d₄) δ 1.06 (s, 9H), 1.39 (s, 9H), 2.39-2.45 (m, 1H),2.65-2.77 (m, 1H), 4.18-4.30 (m, 3H), 4.64 (t, J=8.0 Hz, 1H), 5.72 (m,1H), 6.74 (d, J=8.0 Hz, 1H), 7.44-7.55 (m, 4H), 7.75 (t, J=8.0 Hz, 1H),8.07 (d, J=7.5 Hz, 2H).

LC/MS rt-min (MNa⁺): 2.72 (521) (method B).

Step 2:

Compound 436 was prepared according to Example 420, Step 6 in 66% yield,starting from the product of Example 436, Step 1.

¹H NMR: (DMSO-d₆) δ 0.96 (s, 9H), 1.04-1.05 (m, 2H), 1.09 (m, 2H), 1.30(s, 9H), 1.36-1.39 (m, 1H), 1.71 (t, J=7.5 Hz, 1H), 2.16-2.24 (m, 2H),2.41-2.45 (m, 1H), 2.93 (m, 1H), 4.06-4.09 (m, 3H), 4.38 (br t, 1H),5.10 (d, J=10.5 Hz, 1H), 5.30 (d, J=17 Hz, 1H), 5.60-5.67 (m, 1H), 5.80(m, 1H), 6.49 (br s, 1H), 6.75 (d, J=7.5 Hz, 1H), 7.43-7.51 (m, 3H),7.59 (d, J=7.5 Hz, 1H), 7.81 (t, J=7.5 Hz, 1H), 8.09 (d, J=7.0 Hz, 2H),8.91 (s, NH), 10.4 (s, NH).

LC/MS rt-min (MH⁺): 2.77 (711) (method B).

Example 437 Preparation of Compound 437

Step 1:

To a solution of 2-bromo-6-methyl-pyridine (7.65 g, 44.4 mmol) indichloromethane (50 mL) was added a solution of mCPBA (77%, 12.9 g, 57.7mmol) in dichloromethane (100 mL). The solution was stirred for 18 h atambient temperature. The mixture was neutralized with solid Na₂CO₃ andwater was added. The aqueous phase was extracted with dichloromethane(2×). The combined organic fractions were washed with 5% Na₂S₂O₃, 5%Na₂CO₃, brine and dried (MgSO₄). Purification using a Biotage 40 Mcolumn (eluted gradient hexane-ethyl acetate 40-70%) afforded the titlecompound (5.0 g, 60%) as a colorless oil that solidified upon standing.

¹H NMR: (DMSO-d₆) δ 2.43 (s, 3H), 7.15 (t, J=8.0 Hz, 1H), 7.50 (s, 1H),7.78 (s, 1H).

LC/MS rt-min (MH⁺): 0.32 (188, 190) (method B).

Step 2:

To a solution of the product of Example 437, Step 1 (4.5 g, 24 mmol) inDMF (20 mL) was added POBr₃ (8.2 g, 29 mmol) in portions. A strongexothermic reaction occurred and a precipitate formed. The mixture wasleft for 2 h at ambient temperature. The mixture was quenched with waterand satd. NaHCO₃ until neutral pH. The aqueous phase was extracted withdiethyl ether (2×). The combined organic fractions were washed withbrine and dried (MgSO₄). Purification using a Biotage 40 M column(eluted gradient hexane-diethyl ether 0-10%) afforded the title compound(1.78 g, 30%) as a colorless oil.

¹H NMR: (DMSO-d₆) δ 2.45 (s, 3H), 7.64 (s, 1H), 7.80 (s, 1H).

LC/MS rt-min (MH⁺): 1.85 (250, 252, 254) (method B).

Step 3:

This product was prepared according to Example 421, Step 2 in 46% yield,starting from the product of Example 437, Step 1.

¹H NMR: (DMSO-d₆) δ 1.34, 1.38 (s, 9H (rotamers)), 2.20-2.27 (m, 1H),2.38 (s, 3H), 2.43-2.50 (m, 1H), 3.55 (d, J=12.5 Hz, 1H), 3.64-3.67 (m,1H), 3.66, 3.69 (s, 3H (rotamers)), 4.26-4.32 (m, 1H), 5.17 (m, 1H),6.93 (s, 1H), 7.08 (s, 1H).

LC/MS rt-min (MH⁺): 2.17 (415, 417) (method B).

Step 4:

This product was prepared according to Example 420, Step 3 inquantitative yield, starting from the product of Example 437, Step 3.

This product was prepared according to Example 420, Step 3 in 75% yield,starting from the product of Example 437, Step 3.

¹H NMR: (DMSO-d₆) δ 0.94 (s, 9H), 1.27 (s, 9H), 2.16-2.21 (m, 1H), 2.36(s, 3H), 2.48 (m, 1H (hidden)), 3.64 (s, 3H), 3.83 (d, J=10.5 Hz, 1H),4.06 (d, J=8.5 Hz, 1H), 4.14 (d, J=10.5 Hz, 1H), 4.42 (t, J=9.0 Hz, 1H),5.27 (m, 1H), 6.68 (br s, NH), 6.88 (s, 1H), 7.03 (s, 1H).

LC/MS rt-min (MH⁺): 2.19 (528, 530) (method B).

Step 5:

This product was prepared according to Example 420, Step 5 inquantitative yield, starting from the product of Example 437, Step 4.

¹H NMR: (DMSO-d₆) δ 0.95 (s, 9H), 1.27 (s, 9H), 2.15-2.19 (m, 1H), 2.36(s, 3H), 2.44-2.48 (m, 1H), 3.81 (d, J=10.5 Hz, 1H), 4.04-4.06 (m, 1H),4.13 (d, J=10.5 Hz, 1H), 4.33 (t, J=8.5 Hz, 1H), 5.26 (m, 1H), 6.66 (brd, J=9.0 Hz, NH), 6.88 (s, 1H), 7.03 (s, 1H).

LC/MS rt-min (MNa⁺): 2.14 (536, 538) (method B).

Step 6:

To a solution of the product of Example 437, Step 5 (101 mg, 0.196mmol), Pd(PPh₃)₄ (11.3 mg, 0.0098 mmol), and phenyl boronic acid (34 mg,0.275 mmol) in DMF (2 mL) was added 2M aqueous Na₂CO₃ (0.294 mL, 0.588mmol). The tube was sealed and heated in a microwave oven (Emrys,Personal Chemistry) at 150° C. for 15 min. under nitrogen. After coolingto room temperature the mixture was acidified with 1N HCl (0.5 mL). Thesolid was removed by filtration, and the filtrate concentrated in vacuo.The residue was purified by preparative HPLC (gradient 20-80% B). Thecombined fractions were neutralized with conc. ammonia and concentrated.The residue was partitioned between buffer pH 4 and ethyl acetate. Theaqueous phase was extracted with ethyl acetate (2×) and the combinedorganic extracts washed with brine and dried (MgSO₄). The title compound(115 mg, >100%) was obtained as a white solid.

¹H NMR: (methanol-d₄) δ 1.06 (s, 9H), 1.30 (s, 9H), 2.36-2.41 (m, 1H),2.69 (s, 3H), 2.65-2.76 (m, 1H), 4.01 (dd, J=3.0, 12 Hz, 1H), 4.21 (s,1H), 4.44 (d, J=12 Hz, 1H), 4.64 (dd, J=8.0, 10 Hz, 1H), 5.46 (m, 1H),6.91 (s, 1H), 7.29 (s, 1H), 7.39 (m, 3H), 7.57 (d, J=7.5 Hz, 2H).

LC/MS rt-min (MH⁺): 1.84 (513) (method B).

Step 7:

Compound 437 was prepared according to Example 420, Step 6 in 31% yield,starting from the product of Example 437, Step 6.

¹H NMR: (DMSO-d₆) δ 0.93 (s, 9H), 0.99-1.04 (m, 4H), 1.25 (s, 9H),1.34-1.37 (m, 1H), 1.69-1.71 (m, 1H), 2.11-2.20 (m, 2H), 2.39-2.43 (m,1H), 2.48 (s, 3H), 2.93 (m, 1H), 3.92 (d, J=8.5 Hz, 1H), 4.07 (d, J=9.0Hz, 1H), 4.11 (d, J=12 Hz, 1H), 4.32 (t, J=7.0 Hz, 1H), 5.10 (d, J=10.5Hz, 1H), 5.23 (d, J=17.5 Hz, 1H), 5.39 (m, 1H), 5.60-5.67 (m, 1H), 6.58(d, J=8.5 Hz, 1H), 6.83 (s, 1H), 7.27 (s, 1H), 7.40-7.48 (m, 3H),8.04-8.06 (m, 2H), 8.92 (s, NH), 10.4 (s, NH).

LC/MS rt-min (MH⁺): 2.10 (725) (method B).

Example 438 Preparation of Compound 438

Step 1:

This product was prepared according to Example 437, Step 6 inquantitative yield, starting from 2-thiopheneboronic acid.

¹H NMR: (methanol-d₄) δ 1.05 (s, 9H), 1.32 (s, 9H), 2.32-2.38 (m, 1H),2.56 (s, 3H), 2.66-2.70 (m, 1H), 3.99 (dd, J=3.0, 12 Hz, 1H), 4.23 (s,1H), 4.36 (d, J=12 Hz, 1H), 4.61 (t, J=8.5 Hz, 1H), 5.35 (m, 1H), 5.86(s, 1H), 7.16-7.18 (m, 1H), 7.21 (s, 1H), 7.56 (m, 1H), 7.72 (d, J=3.0Hz, 1H).

LC/MS rt-min (MH⁺): 1.80 (519) (method B).

Step 2:

Compound 438 was prepared according to Example 420, Step 6 in 41% yield,starting from the product of Example 438, Step 1.

¹H NMR: (DMSO-d₆) δ 0.91 (s, 9H), 0.99-1.04 (m, 4H), 1.20 (s, 9H),1.35-1.37 (m, 1H), 1.69-1.71 (m, 1H), 2.09-2.20 (m, 2H), 2.33 (m, 1H),2.42 (s, 3H), 2.93 (m, 1H), 3.92 (d, J=8.5 Hz, 1H), 4.07-4.11 (m, 2H),4.29-4.32 (m, 1H), 5.10 (d, J=10.5 Hz, 1H), 5.23 (d, J=17.0 Hz, 1H),5.36 (m, 1H), 5.60-5.67 (m, 1H), 6.58 (d, J=8.5 Hz, 1H), 6.75 (s, 1H),7.14 (t, J=4.5 Hz, 1H), 7.28 (s, 1H), 7.59 (d, J=5.5 Hz, 1H), 7.80 (d,J=3.5 Hz, 1H), 8.92 (s, NH), 10.4 (s, NH).

LC/MS rt-min (MH⁺): 2.06 (731) (method B).

Section K Example 450 Preparation of Compound 450

Compound 450 was prepared according to Example 8, Step 5, except using4-chloro-6-fluoro-2-trifluoromethylquinoline instead.

¹H NMR (CD₃OD) δ 0.97-1.04 (m, 12H), 1.17-1.24 (m, 10H), 1.39-1.46 (m,1H), 1.82-1.87 (m, 1H), 2.20-2.23 (m, 1H), 2.35-2.39 9m, 1H), 2.55-2.65(m, 1H), 2.91-2.96 (m, 1H), 4.09-4.11 (m, 1H), 4.18-4.21 (m, 1H), 4.56(b, 2H), 5.10-5.14 (m, 1H), 5.28-5.31 (m, 1H), 5.60 (b, 1H), 5.70-5.80(m, 1H), 7.41 (s, 1H), 7.65-7.68 (m, 1H), 7.86 (s, 1H), 8.13-8.15 (m,1H).

Example 451 Preparation of Compound 451

Step 1:

The tosylate was prepared as described in the literature (Patchett, A.A.; Witkof, B. J. Am. Chem. Soc. 1957, 185-192) and was used withoutfurther purification.

To a slurry of NaH (76 mg, 1.90 mmol) in DMF (20 ml) was added1-thionaphthol (0.29 mg, 1.80 mmol) and the mixture stirred for 30minutes. A solution of the tosylate (0.61 g, 1.80 mmol) was added andthe mixture stirred for 12 h at 23° C. The mixture was concentrated andthe residue partitioned between EtOAc/H₂O. The organic extracts aredried (MgSO₄) and concentrated. The residue was purified by columnchromatography (elution with 5% EtOAc/hexanes to 30% EtOAc/hexanes togive 261 mg (38%) of the product as a yellow oil.

¹H NMR (CDCl₃, 3:2 mixture of rotamers) δ 1.41 (s, 9H), 1.44 (s, 9H),2.25-2.29 (m, 2H), 3.69 (s, 3H), 3.35-3.42 (m, 1H), 3.51-3.53 (m, 1H),3.80-3.86 (m, 2H), 4.38-4.39 (m, 1H), 4.46-4.48 (m, 1H), 7.41-7.46 (m,1H), 7.42-7.54 (m, 1H), 7.57-7.59 (m, 1H), 7.58 (d, J=4 Hz, 1H),7.82-7.88 (m, 2H), 8.46 (d, J=5 Hz, 1H);

LC-MS (retention time: 1.93), MS m/z 388 (M⁺+1).

Step 2:

A mixture of 4-(naphthalen-1-ylsulfanyl)-pyrrolidine-1,2-dicarboxylicacid 1-tert-butyl ester 2-methyl ester (0.38 g, 0.98 mmol) and 4N HCl(1.0 ml) was stirred at 23° C. for 2 h. The solvent was removed and theresidue dissolved in CH₃CN (20 ml) and treated with acid (0.37 g, 2.16mmol), TBTU (0.23 g, 0.98 mmol) and DIPEA (0.37 g, 2.16 mmol) andstirred for 12 h. The mixture is concentrated and the residue dissolvedin EtOAc and washed with 1 N HCl, saturated NaHCO₃ then dried over MgSO₄and concentrated. The residue was used without further purification.

¹H NMR (CDCl₃, 1:1 mixture of rotamers) δ 0.99 (s, 9H), 1.02 (s, 9H),1.44 (s, 9H), 1.46 (s, 9H) 2.2-2.25 (m, 2H), 3.70 (s, 3H), 3.82-3.86 (m,1H), 3.89-3.92 (m, 2H) 4.26 (s, 1H), 4.28 (s, 1H), 4.70-4.75 (m, 1H),7.40-7.48 (m, 1H), 7.54-7.55 (m, 1H), 7.59-7.62 (m, 1H), 7.72-7.74 (m,1H), 7.86-7.89 (m, 2H), 8.48-8.50 (m, 1H);

LC-MS (retention time: 1.59), MS m/z 523 (M+Na).

Step 3:

To a mixture of1-(2-tert-butoxycarbonylamino-3,3-dimethyl-butyryl)-4-(naphthalen-1-ylsulfanyl)-pyrrolidine-2-carboxylicacid methyl ester (Example 451, Step 2) (0.49 g, 0.98 mmol), in THF/H₂O(2:1) was added LiOH hydrate (0.20 g, 4.9 mmol) and the mixture stirredfor 12 h. The solution was concentrated and washed with EtOAc. Theaqueous layer is acidified with 1N HCl and extracted with EtOAc. Theproduct was observed in the first EtOAc extract. The first organicextract was dried over MgSO₄ and concentrated to 328 mg (71%) of a tansolid.

¹H NMR (DMSO-d₆, 2:1 mixture of rotamers) δ 0.88 (s, 9H), 0.92 (s, 9H),1.34 (s, 9H), 1.38 (s, 9H), 2.18-2.25 (m, 2H), 3.66-3.75 (m, 1H),3.89-4.00 (m, 2H), 4.10-4.13 (m, 1H), 4.25-4.32 (m, 1H), 7.45-7.7.51 (m,1H), 7.56-7.61 (m, 2H), 7.65-7.7.69 (m, 1H), 7.88-7.91 (m, 1H), 7.97 (d,J=4.8 Hz, 1H), 8.27-8.35 (m, 1H);

LC-MS (retention time: 1.52), MS m/z 486 (M+1).

Step 4:

To a solution of the acid (Example 451, Step 3) (0.32 g, 0.87 mmol) inCH₃CN (10 mL) and DMF (2 mL) was added the diastereomeric mixture of1(R)-2(S) and 1-(S)-2(R) 1-amino-2-vinyl-cyclopropanecarboxylic acidethyl ester hydrochloride (240 mg, 0.87 mmol) and TBTU (201 mg, 0.87mmol) and DIPEA (0.32 mL, 0.742 mmol) and the mixture stirred at 23° C.for 12 h. The mixture was concentrated and the residue partitionedbetween EtOAc and water. The organic layer was separated, dried overMgSO₄ and concentrated. The residue was chromatographed with 30%EtOAc/hexanes as eluant to give 240 mg (38%) of a light yellow solid.

¹H NMR (DMSO-d₆, mixture of rotamers and diasteromers) δ 0.87 (s, 9H),0.88 (s, 9H), 0.97-1.04 (m, 3H), 1.33 (s, 9H), 1.38 (s, 9H), 2.11-2.20(m, 2H), 3.74-3.85 (m, 1H), 3.89-3.96 (m, 2H), 3.98-4.03 (m, 4H),4.00-4.09 (m, 1H), 4.40-4.42 (m, 1H), 5.04-5.09 (m, 1H), 5.17-5.29 (m,2H), 5.50-5.70 (m, 1H), 6.60-6.62 (m, 1H), 6.72-6.75 (m, 1H), 7.50-7.56(m, 1H), 7.56-7.72 (m, 2H), 7.70-7.80 (m, 1H), 7.92-8.00 (m, 1H),8.00-8.06 (m, 1H), 8.29-8.40 (m, 1H), 8.65 (s, 1H), 8.79 (s, 1H);

LC-MS (retention time: 1.59), MS m/z 623 (M+1).

Step 5:

The acid was prepared as previously described using LiOH in THF/MeOH/H₂O(4/2/1) in Example 451, Step 3, except using the product of Example 451,Step 4 instead.

¹H NMR (DMSO-d₆, mixture of rotamers and diastereomers) δ 0.90 (s, 9H),1.17-1.23 (m, 2H), 1.32-1.37 (m, 9H), 2.10-2.12 (m, 1H), 2.20-2.31 (m,2H), 3.97-4.05 (m, 2H), 4.10-4.12 (m, 1H), 4.32-4.40 (m, 1H), 4.55-4.61(m, 1H), 4.80-4.98 (m, 2H), 5.03-5.08 (m, 1H), 5.10-5.20 (m, 1H),5.75-5.90 (m, 1H), 6.55-6.70 (m, 1H), 7.42-7.57 (m, 1H), 7.60-7.64 (m,2H), 7.70-7.72 (m, 1H), 7.80-7.97 (m, 1H), 7.96-7.99 (m, 1H), 8.20-8.50(m, 2H);

LC-MS (retention time: 1.52), MS m/z 595 (M+1).

Step 6:

A mixture of the acid (Example 451, Step 5) (172 mg, 0.29 mmol),methanesulfonamide (110 mg, 1.16 mmol), EDAC (110 mg, 0.58 mmol) andDMAP (71 mg, 0.58 mmol) was dissolved in THF (10 ml) and stirred for 12h. DBU (0.087 mL, 0.58 mmol) was added and the mixture stirred for 48 h.The solvent is removed and the residue dissolved in EtOAc and washedwith water and 1N HCl, dried over MgSO₄ and concentrated. The residuewas purified by preparative thin layer chromatography to give 15 mg (8%)of Compound 451 as a tan solid.

¹H NMR (DMSO-d₆, mixture of rotamers and diastereomers) δ 0.98 (s, 9H),1.27-1.42 (m, 2H), 1.46 (s, 9H), 1.76-1.79 (m, 1H), 1.83-1.86 (m, 1H),1.92-2.10 (m, 1H), 2.16-2.25 (m, 2H), 3.01-3.10 (m, 1H), 3.80-3.83 (m,1H), 3.86-3.89 (m, 1H), 3.98-3.99 (m, 1H), 3.99-4.05 (m, 1H), 4.24-4.29(m, 1H), 4.44-4.53 (m, 1H), 4.86 (s, 3H), 5.08-5.15 (m, 1H), 5.25-5.29(m, 1H), 5.65-5.85 (m, 1H), 6.5-6.8 (m, 1H), 7.48 (t, J=7.7 Hz, 1H),7.54 (t, J=7.1 Hz, 1H), 7.59 (t, J=7.1 Hz, 1H), 7.75-7.77 (m, 1H),7.88-7.91 (m, 2H), 8.46 (d, J=8.25 Hz, 1H);

LC-MS (retention time: 1.52), MS m/z 672 (M+1 minor), m/z 693 (M+NaMajor).

Example 452 Preparation of Compound 452

Step 1:

To a slurry of NaH (76 mg, 1.90 mmol) in DMF (20 mL) is added2-thionaphthol (0.29 g, 1.80 mmol) and the mixture is stirred for 30minutes. A solution of the tosylate (Example 451, Step 1) (0.61 g, 1.79mmol) in DMF (2 ml) is added and the mixture stirred for 12 h ar 23° C.The mixture is concentrated then partitioned between EtOAc/H₂O. Theorganic layer was washed with saturated NaHCO₃, dried (MgSO₄) andconcentrated. The residue was chromatographed with 5% EtOAc/hexanesfollowed by 30% EtOAc/hexanes to give 261 mg (38%) of the product as aclear oil.

¹H NMR (DMSO-d₆) δ 1.32 (s, 9H), 2.29-2.35 (m, 2H), 3.33-3.47 (m, 2H),3.66 (s, 3H), 3.71-3.81 (m, 1H), 4.29-4.32 (s, 1H), 7.49-7.55 (m, 3H),7.70-7.80 (m, 1H), 7.81-7.97 (m, 3H);

LC-MS (retention time: 1.54), MS m/z 387 (M+1).

Step 2:

A mixture of 4-(naphthalen-2-ylsulfanyl)-pyrrolidine-1,2-dicarboxylicacid 1-tert-butyl ester 2-methyl ester (310 mg, 0.80 mmol) and 4N HCl indioxane (1.49 ml, 2.69 mmol) was stirred for 2 h at 23° C. thenconcentrated. The residue is dissolved in CH₃CN (10 mL) andN-Boc-t-butylglycine (196 mg, 0.85 mmol), TBTU (0.27 g, 0.85 mmol) andDIPEA (0.32 mL, 1.85 mmol) were added and the mixture stirred overnight.The mixture was concentrated and the residue dissolved in EtOAc, washedwith 1 N HCl, saturated NaHCO₃, dried and concentrated to give 300 mg(90%) of the product as a yellow oil.

¹H NMR (Methanol-d₄) δ 0.99 (s, 9H), 1.44 (s, 9H), 2.20-2.35 (m, 2H),3.75 (s, 3H), 3.92-4.08 (m, 2H), 4.26 (d, J=9.4 Hz, 1H), 4.57 (t, J=9.5Hz, 1H) 6.46 (d, J=9.5 Hz, 1H), 7.48-7.60 (m, 3H), 7.83-7.90 (m, 3H),8.02 (s, 1H)

LC-MS (retention time: 1.98), MS m/z 523 (M+Na).

Step 3:

A solution of 4-(naphthalen-2-ylsulfanyl)-pyrrolidine-1,2-dicarboxylicacid 1-tert-butyl ester 2-methyl ester (0.48 g, 0.96 mmol) is dissolvedin MeOH (20 mL) and stirred with LiOH (0.2 g, 4.8 mmol) for 12 h. thesolution is concentrated and acidified and extracted with EtOAc. Theorganic extract was dried over MgSO₄ and concentrated to give 418 mg(91%) of a yellow solid.

¹H NMR (DMSO-d₆, 1:2 mixture of rotamers) δ 0.86, 0.93 (s, 9H) (1:2mixture of rotamers), 1.35, 1.38 (s, 9H) (1:2 mixture of rotamers),2.01-2.18, 2.25-2.35 (m, 2H), 3.25-3.40 (m, 2H), 3.70-3.80 (m, 1H),4.00-4.20 (m, J=9.4 Hz, 2H), 4.30-4.40 (s, 1H), 5.61-5.70, 6.42-6.50 (m,1H) (1:2 mixture of rotamers), 7.50-7.54 (m, 3H), 7.87-7.89 (m, 3H),7.98 (s, 1H);

LC-MS (retention time: 1.93), MS m/z 487 (M+1).

Step 4:

A solution of 1-tert-butoxycarbonylamino-2-vinyl-cyclopropanecarboxylicacid ethyl ester (4.3 g, 17.8 mmol) in MeOH (50 mL) was treated withLiOH (0.84 g, 20.0 mmol) and water (5 mL) and the mixture stirred for 12h. The solvent was removed and the residue acidified and extracted withEtOAc. The organic layers are dried over MgSO₄, filtered andconcentrated to give the acid 2.1 g (52%) as a yellow oil. The acid (2.1g, 9.25 mmol) was dissolved in THF and treated with CDI (7.25 g, 13.8mmol) and heated to reflux for 3 h then cooled to 23° C.Methanesulfonamide (1.76 g, 18.5 mmol) was added followed by DBU (2.77ml, 18.5 mmol) and stirred for 72 h at 23° C. The mixture wasconcentrated and the residue acidified to pH 4 (1 N HCl) and extractedwith EtOAc. The organic extracts were dried over MgSO₄ and concentratedto give 1.99 g (71%) of a yellow oil that solidified on standing.

¹H NMR (DMSO-d₆) δ 1.16-1.23 (m, 1H), 1.43 (s, 9H), 1.65-1.75 (m, 1H),2.15-2.25 (m, 2H), 3.16 (s, 3H), 5.08 (d, J=9.9 Hz, 1H), 5.22 (d, J=17.1Hz, 1H), 5.40-5.52 (m, 1H);

LC-MS (retention time: 1.14), MS m/z 304 (M+1).

Step 5:

A solution of the product of Example 452, Step 4 in dioxane/4N HCl (2ml) was stirred for 2 h then concentrated. The residue was dissolved inCH₃CN (5 mL) and added to a mixture of the acid (Example 452, Step 3)(120 mg, 0.25 mmol), TBTU (58 mg, 0.25 mmol) and DIPEA (0.06 ml, 0.35mmol) was added and the mixture stirred for 12 h. The solvent wasremoved and the residue dissolved in EtOAc and washed with 1N HCl,saturated NaHCO₃ dried over MgSO₄ and concentrated. The residue ispurified with preparative TLC (Analtech 20×40 cM, 1000 SiO₂) to give 128mg (70%) of Compound 452 as a tan solid.

¹H NMR (DMSO-d₆, mixture of diasteromers) δ 0.97, 0.99 (s, 9H),1.23-1.43 (m, 2H), 1.45 (s, 9H), 1.82-1.83 (m, 1H), 2.00-2.51 (m, 2H),2.90-2.99 (m, 1H), 3.33 (s, 3H), 3.90-3.99 (m, 1H), 4.01-4.20 (m, 2H),4.25-4.30 (m, 1H), 4.45-4.55 (m, 1H), 4.95-5.10 (m, 1H), 5.12-5.25 (m,1H), 5.71-5.85 (m, 1H), 6.4-6.8 (br m, 1H), 7.46-7.53 (m, 3H), 7.80-7.86(m, 3H), 7.98-7.99 (m, 1H);

LC-MS (retention time: 1.95), MS m/z 672 (M+1).

Example 453 Preparation of Compound 453

Step 1:

A mixture of Example 452, Step 3 (110 mg, 0.23 mmol) in DCM (20 ml),3-chloroperbenzoic acid (121.6 mg, 0.57 mmol, 85% peracid), KHPO₄ (0.13g, 0.94 mmol) and K₂HPO₄ (0.18 g, 1.05 mmol) are stirred at 23° C. for12 h. The solution is diluted with DCM, washed with water, saturatedNaHCO₃, dried over MgSO₄ and concentrated to give the product 110 mg,(92%) as a clear oil.

¹H NMR (DMSO-d₆) δ 0.91 (s, 9H), 1.48 (s, 9H), 2.23-2.28 (m, 1H),2.65-2.80 (m, 1H), 3.88-3.90 (m, 1H), 4.12 (t, J=8.0 Hz, 1H), 4.20 (d,J=9.5 Hz, 1H), 4.27 (d, J=9.9 Hz, 1H), 6.76 (d, J=9.3 Hz, 1H), 7.70-7.80(m, 2H), 7.88-7.95 (m, 1H), 8.11 (d, J=8.1 Hz, 1H), 8.17 (d, J=8.6 Hz,1H), 8.26 (d, J=8.1 Hz, 1H), 8.71 (s, 1H);

LC-MS (retention time: 1.72), MS m/z 519 (M+1).

Step 2:

A mixture of the product of Example 453, Step 1 (110 mg, 0.212 mmol),amine (Example 452, Step 5a) (0.65 mg, 0.212 mmol), TBTU (48.5 mg, 0.21mmol) followed by DIPEA (60.8 ml, 0.35 mmol) and stirred for 12 h at 23°C. The solvent is removed and the residue dissolved in EtOAc and washedwith 1N HCl, saturated NaHCO₃, dried over MgSO₄ and concentrated. Theresidue was purified by preparative TLC (eluted with 10% MeOH/CH₂Cl₂) togive 25 mg (17%) of Compound 453 as a white solid.

¹H NMR (DMSO-d₆, mixture of diastereomers) δ 0.93, 0.96 (s, 9H),1.38-1.45 (m, 2H), 1.53, 1.55 (m, 9H), 1.76-1.85 (m, 1H), 2.21-2.40 (m,2H), 3.13-3.15 (m, 2H), 3.34 (s, 3H), 3.91-3.99 (m, 1H), 4.15 (m, 1H),4.25 (m, 1H), 4.30 (m, 1H), 5.09-5.12 (m, 1H), 5.26-5.31 (m, 1H),5.72-5.76 (m, 1H), 6.65-6.68 (m, 1H), 6.71-6.76 (m, 1H), 7.67-7.76 (m,2H), 7.91-7.95 (m, 1H), 8.03 (d, J=7.8 Hz, 1H), 8.13 (d, J=8.5 Hz, 1H),8.20 (d, J=7.6 Hz, 1H), 8.7 (s, 1H);

LC-MS (retention time: 1.76), MS m/z 705 (M+1).

Example 454 Preparation of Compound 454

Step 1:

To slurry of the sodium hydride (0.91 g, 22.7 mmol) in THF (50 mL) wasadded N—BOC-trans-4(R)-hydroxy-L-proline (2.5 g, 10.8 mmol) and themixture stirred at 23° C. for 1 h. 2-Chloromethylnapthalene (1.9 g, 10.8mmol) was added and the mixture stirred for 12 h at room temperature.The solvent was removed and the residue poured into water and washedwith hexanes. The aqueous layer was acidified (1 N HCl) and extractedwith EtOAc. The EtOAc layer is separated, dried (MgSO₄), andconcentrated to give a light yellow residue. The oil was purified byflash chromatography with 1:1 EtOAc/hexanes with 1% acetic acid added togive 1.56 g (39%) of the desired product as a thick oil.

¹H NMR (DMSO-d₆, 3:1 mixture of rotamers) δ 1.35, 1.37 (s, 9H, major andminor respectively), 1.92-2.02, 2.15-2.20 (m, 2H, major and minorrespectively), 2.35-2.50 (m, 2H), 3.41-3.49 (m, 2H), 4.12-4.16,4.20-4.21 (m, 2H), 4.65-4.68 (m, 2H), 7.46-7.52 (m, 3H), 7.74-7.91 (m,4H), (Acid OH not observed);

LC-MS (retention time: 1.44, YMC ODS-A C18 S7 3.0×50 mm, gradient 10%MeOH/H₂O 0.1% TFA to 90% MeOH/H₂O 0.1% TFA), MS m/z 394 (M++1+Na).

Step 2:

To a solution of the HCl salt of a 1:1 mixture of diastereoisomers(1R,2S/1S,2R where carboxy group is syn to vinyl moiety) of2-(1-ethoxycarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-(naphthalen-2-ylmethoxy)-pyrrolidine-1-carboxylicacid tert-butyl ester (0.54 g, 1.3 mmole)[prepared by stirring the N-Bocamine with HCl (4N) in dioxane for 1 hr then removal of the solvent invacuo] in CH₃CN (50 mL) is added Boc-4(R)-(2-methylnapthyl)proline (0.5g, 1.3 mmol), TBTU (0.45 g, 1.4 mmol) followed by DIPEA (0.78 mL, 4.5mmol). The mixture is stirred for 12 h and concentrated. The residue wasdissolved in EtOAc/H₂O and washed with saturated NaHCO₃, saturated NaCl,dried (MgSO₄) and concentrated to give a thick yellow oil (0.6 g, 91%)of the product as a mixture of diastereomers.

¹H NMR (DMSO-d₆) δ: 1.08-1.22 (m, 7H), 1.23-1.39 (m, 9H), 2.02-2.18 (m,1H), 2.25-2.35 (m, 1H), 3.33-3.53 (m, 2H), 3.90-4.14 (m, 4H), 4.45-4.70(m, 2H), 5.07-5.11 (m, 1H), 5.24-5.30 (m, 1H), 5.58-5.63 (m, 1H),7.43-7.51 (m, 4H), 7.84-7.96 (m, 3H); MS m/z 531 (M⁺+1+Na).

Step 3:

A solution of product of Example 454, Step 2 (600 mg, 1.18 mmol) wasstirred with HCl (4N, 3 mL, 11.8 mmol) in dioxane for 1 hr then thesolvent removed in vacuo. The residue is dissolved in CH₃CN (10 mL)treated with Boc-L-t-Bu-Gly (0.42 g, 1.38 mmol), TBTU (0.27 g, 1.18mmol) followed by DIPEA (0.71 mL, 4.1 mmol). The mixture is stirred for12 h and concentrated. The residue was dissolved in EtOAc/H₂O and washedwith 1N HCl, saturated NaHCO₃, saturated NaCl, dried (MgSO₄) andconcentrated to give a thick yellow oil. The product was purified byflash chromatography using gradient elution 5% EtoAc/Hexanes 10%EtOAc/Hexanes, 30% EtOAc/Hexanes and finally as eluant to give theproduct as a thick oil (0.243 g, 33%) of the product as a mixture ofdiastereomers and rotamers.

¹H NMR (DMSO-d₆) δ: 0.83-1.00 (m, 10H), 1.34 (s, 9H), 1.58-1.59,1.65-1.67 (m, 2H), 1.95-1.99, 2.04-2.06, 2.10-2.19, 2.24-2.56 (m, 2H),3.97-4.04 (m, 3H), 4.08-4.17 (m, 3H), 4.29-4.31 (m, 2H), 4.59-4.72 (m,3H), 5.06-5.10 (m, 1H), 5.18-5.30 (m, 1H), 5.60-5.63 (m, 1H), 6.59-6.65,6.70-6.74 (m, 1H), 7.43-7.51 (m, 4H), 7.84-7.96 (m, 3H), 8.66, 8.76 (s,1H);

MS m/z 531 (M⁺+1+Na).

Step 4:

To a suspension of product of Example 454, Step 3 (240 mg, 0.39 mmol) inTHF (15 mL), and H₂O (2 mL) was added LiOH (82 mg, 1.95 mmol). Thereaction mixture was stirred for 12 h then concentrated in vacuo untilonly the aqueous layer remained. The resulting aqueous residue wasacidified to pH 3.0 by addition of 1.0 N aqueous HCl, and extracted withEtOAc (2×80 mL). Combined organic extracts was dried (MgSO₄), filtered,and concentrated in vacuo to give the product as a tan solid (200 mg,0.33 mmol, 85%):

¹H NMR (DMSO-d₆) δ: 0.86, 0.94 (s, 9H minor and major respectively),1.23-1.42 (m, 2H), 1.34 (s, 9H), 1.8-2.1 (m, 2H), 2.18-2.30 (m, 1H),3.59-3.73 (m, 3H), 4.0-4.09 (m, 1H), 4.18-4.34 (m, 3H), 4.56-4.62 (m,1H), 4.66-4.67 (m, 1H), 4.82-4.92 (m, 1H), 5.0-5.20 (m, 1H), 5.91-6.08(m, 1H), 6.5-6.7 (m, 1H), 7.45-7.59 (m, 3H), 7.82-7.97 (m, 4H), 8.2-8.3,8.3-8.4 (s, 1H);

LC-MS (retention time: 1.50), MS m/z 593 (M⁺+1).

Step 5:

To a solution of product of Example 454, Step 4 (190 mg, 0.32 mmol) andEDAC (122 mg, 0.64 mmol) and 4-DMAP (78 mg, 0.64 mmol) in THF (20 mL)was added commercially available methanesulfonamide (122 mg, 1.28 mmol).The resulting solution was stirred for 2 days, then DBU was added (95μL, 0.64 mmol). The reaction was stirred for 24 h then concentrated. Theresidue was partitioned between EtOAc (80 mL) and water and washed with1 N HCl), aqueous NaHCO₃ (2×30 mL), dried (MgSO₄) and purified bypreparative HPLC (65-90% MeOH/Water/0.1% TFA) which gave 56 mg of amixture of product and material in which the BOC group was removed. Thematerial was further purified by preparative TLC (eluted with 10%MeOH/CH₂Cl₂ using 20×40 cM plates from Analtech) to give Compound 454 asa tan solid (12 mg, 6%).

¹H NMR (MeOD-d₄ 50/50 mixture of P1 diastereomers) δ 0.88-0.99 (m, 2H),1.01, 1.02 (s, 9H minor and major diastereomers respectively), 1.23-1.42(m, 2H), 1.38 (s, 9H), 1.72-1.79 (m, 1H), 1.86-1.88 (m, 1H), 2.00-2.10(m, 2H), 2.10-2.23 (m, 1H), 2.3-2.5 (m, 1H), 3.12, 3.17 (s, 3H),3.72-3.79 (m, 1H), 4.26-4.41 (m, 3H), 4.72 (d, J=8.2 Hz, 1H), 4.76 (d,J=8.2 Hz, 1H), 5.09-5.12 (t, J=9.3 Hz, 1H), 5.28 (dd, J=3.5, 17.6 Hz,1H) 5.7-5.8 (m, 1H), 6.55-6.80 (m, 1H), 7.45-7.47 (m, 3H), 7.79-7.83 (m,4H);

LC-MS (retention time: 1.48), MS m/z 670 (M⁺+1).

Example 470 Preparation of Compound 470

Step 1:

To a solution of commercially availableN-Boc-(4S)-(cis)-Hydroxyproline-OMe (200 mgs, 0.82 mmole),triphenylphosphine (320 mgs, 1.22 mmole) and 1-naphthol (176 mgs, 1.22mmole) in 2.5 mL tetrahydrofuran was added dropwise a solution ofdiethyldiazodicarboxylate (190 μL, 1.22 mmole) in 1.0 mL THF over 10minutes. After stirring for 5.5 days, the reaction was concentrated invacuo. The crude yellow oil was chromatographed on a 20×40 cMpreparative TLC plate (Analtech SiO2) eluting with 6-1 hexanes-ethylacetate to yield the desired product as a pale yellow oil (150 mgs,33%).

¹H NMR (CDCl₃, 500 MHz) δ 1.44 (s, 9H) 2.33 (1H, m), 2.72 (1H, m), 3.77and 3.38 (2s, 3H, rotamers), 3.88 (dd, 1H, J=4.3, 12.4 Hz), 3.97 (bd,1H), 4.53 and 4.62 (2t, 1H, J=7.8 Hz, rotamers), 5.10 (bd, 1H), 6.76 (t,1H, J=9.5 Hz), 7.37 (m, 1H), 7.46 (m, 3H), 7.80 (d, 1H, J=7.7 Hz), 8.18(m, 1H);

LC-MS A (retention time: 1.86; MS m/z 394 (M+Na)⁺

Step 2:

To a stirred solution of Boc-(4R)-naphthal-1-oxo)-Pro-OEt (150 mgs, 0.40mmole) in 1.5 mL THF and 0.5 mL water was added lithium hydroxide (10mgs). The solution was stirred for 21 hours at room temperature and thendiluted with 0.5N NaHCO₃. The basic solution was extracted with ethylacetate and then the aqueous layer was acidified to pH 2 with thedropwise addition of conc. HCl. This acidified layer was then extractedagain with ethyl acetate. This second ethyl acetate layer was dried withmagnesium sulfate, filtered and then concentrated in vacuo to yieldBoc-(4R)-naphthal-1-oxo)-Pro-OH as pale-pink crystals (147 mgs, 100%).

¹H NMR (CDCl₃, 500 MHz) δ 1.47 and 1.48 (2s, 9H, rotamers), 2.40 and2.52 (2m, 1H), 2.68 and 2.78 (2m, 1H), 3.78-4.07 (m, 2H), 4.57 and 4.69(2t, 1H, J=7.6 and 8.0 Hz, rotamers), 5.12 (bd, 1H), 6.77 (dd, 1H,J=7.6, 21.2 Hz), 7.37 (m, 1H), 7.46 (m, 3H), 7.81 (t, 1H, J=5.8 Hz),8.19 (m, 1H);

LC-MS A (retention time: 1.79; MS m/z 358 (M+H)⁺

Step 3:

To a solution of Boc-((4R)-naphthal-1-oxo)-Pro-OH (147 mgs, 0.41 mmole)and racemic(1R/2S)/(1S/2R)-1-amino-2-vinylcyclopropane carboxylic acidethyl ester hydrochloride salt (79 mgs, 0.41 mmole) in 2.8 mL methylenechloride was added DIPEA (250 μL, 1.44 mmole) and TBTU (158 mgs, 0.49mmole). The resulting solution was stirred under nitrogen for 20 hoursand then diluted with 40 mL methylene chloride. The organic layer waswashed with water, 1N NaHCO₃, 1N HCl, water and brine. The solution wasthen dried with sodium sulfate and concentrated in vacuo. Purificationby preparative TLC yielded two separate diastereomers, higher Rfdiastereomer A (P2[Boc(4R)-(naphthal-1-oxo)proline]-P1(1R,2S VinylAcca)-OEt, 78 mgs, 38%) and lower Rf diastereomer B(P2[Boc(4R)-(naphthal-1-oxo)proline]-P1(1S,2R Vinyl Acca)-OEt, 91 mgs,45%) as off white solids:

Diastereomer A: P2[Boc(4R)-(naphthal-1-oxo)proline]-P1(1R,2S VinylAcca)-OEt:

¹H NMR (CDCl₃, 500 MHz) δ 1.24 (t, 3H), 1.43 (s, 9H), 1.52 (m, 1H), 1.84(m, 1H), 2.02 (m, 1H), 2.14 (m, 1H), 2.81 (m, 1H), 3.88 (m, 2H), 4.11(q, 1H, J=7.15), 4.19 (m, 1H), 4.54 (m, 1H), 5.15 (m, 1H), 5.31 (dd, 1H,J=17, 0.8 Hz), 5.77 (m, 1H), 6.83 (m, 1H), 7.36 (t, 1H, J=7.8 Hz), 7.46(m, 3H), 7.78 (d, 1H, J=7.6 Hz), 8.14 (d, 1H, J=8.15 Hz);

LC-MS B (retention time: 1.85; MS m/z 495 (M+H)⁺

Diastereomer B, Example 10B:P2[Boc(4R)-(naphthal-1-oxo)proline]-P1(1S,2R Vinyl Acca)-OEt: ¹H NMR(dl-CHCl₃, 500 MHz) δ 1.24 (t, 3H), 1.42 (s, 9H), 1.85 (m, 1H), 2.15 (q,1H, J=8.9 Hz), 2.40 (m, 1H), 2.78 (m, 1H), 3.78 (m, 1H), 4.12 (m, 2H),4.52 (m, 1H), 5.15 (m, 1H), 5.31 (m, 1H), 5.79 (m, 1H), 6.80 (m, 1H),7.35 (t, 1H, J=7.6 Hz), 7.46 (m, 3H), 7.78 (d, 1H, J=7.6 Hz), 8.14 (d,1H, J=8.10 Hz).

LC-MS B (retention time: 1.85; MS m/z 495 (M+H)⁺

Step 4:

To P2[Boc(4R)-(naphthal-1-oxo)proline]-P1(1R,2S Vinyl Acca)-OEt (A,higher Rf) (78 mg, 0.16 mmol) was added 4N HCl in dioxane (2.0 mL) andthe solution was allowed to stir for 30 minutes. Concentration in vacuoyielded the HCl salt of P2[(4R)-(naphthal-4-oxo) proline)]-P1(1R,2SVinyl Acca)-OEt as a yellow oil which was taken on to the next stepdirectly without further purification. To a solution of BOC L-tBuGly (73mgs, 0.32 mmole) and the HCl salt of P2[(4R)-(naphthal-4-oxo)proline)]-P1(1R,2S Vinyl Acca)-OEt (0.16 mmole) in 11 mL acetonitrilewas added DIPEA (140 μL, 0.79 mmole) and HATU (132 mgs, 0.35 mmole). Theresulting solution was stirred under nitrogen for 17 hours and thendiluted with 100 mL ethyl acetate. The organic layer was washed withwater, 1N NaHCO₃, 1N HCl, water and brine. The solution was then driedwith sodium sulfate and concentrated in vacuo to yield the titlecompound as a pale-yellow oily film (92 mgs, 96%).

¹H NMR (CDCl₃, 500 MHz) δ 1.06 (s, 9H), 1.22 (t, 3H, J=7.1), 1.38 (s,9H), 1.41 (m, 1H), 1.82 (m, 1H), 2.13 (m, 1H), 2.42 (m, 1H), 2.79 (m,1H), 3.92-4.2 (m, 1H), 4.12 (q, 2H, J=6.6 Hz), 4.38 (bt, 1H), 5.12 (d,1H, J=10.3 Hz) 5.2-5.39 (m, 3H), 5.75 (m, 1H), 6.82 (d, 1H, J=7.5 Hz),7.34-7.46 (m, 4H), 7.59 (bs, 1H, NH), 7.76 (d, 1H, J=7.9 Hz), 8.13 (d,1H, J=8.3 Hz);

LC-MS C (retention time: 2.82; MS m/z 608 (M+H)⁺

Step 5:

To a solution of product of Example 470, Step 4 (92 mgs, 0.15 mmole) in750 mL tetrahydrofuran and 250 mL water was added lithium hydroxide (4mgs). The resulting solution was stirred for 28.5 hours worked up asusual and then resubjected to the same conditions except adding twice asmuch lithium hydroxide (8 mgs). After 24 hours the reaction was dilutedwith ethyl acetate and washed with water. The organic layer was driedwith sodium sulfate and concentrated in vacuo. The resulting semisolidwas purified by flash chromatography eluting with 3-1 hexanes-ethylacetate to yield BocNH-P3(t-BuGly)-P2[(Boc (4R)-(naphthal-1-oxo)proline)]-P1(1R,2S Vinyl Acca)-OH as a clear semisolid (30 mgs, 34%).

¹H NMR (d₄-MeOH, 500 MHz) δ 1.04 (s, 9H), 1.24 (t, 1H, J=3.9 Hz), 1.32(s, 9H), 1.66 (m, 1H), 2.07 (m, 1H), 2.40 (m, 1H), 2.71 (m, 1H),4.04-4.07 (m, 1H), 4.28 (m, 1H), 4.42 (m, 1H), 4.55 (m, 1H), 5.02 (m,1H), 5.18-5.29 (m, 2H), 5.90 (m, 1H), 6.54 (m, 1H), 6.92 (m, 1H), 4.26(m, 4H), 7.77 (m, 1H), 8.15 (m, 1H);

LC-MS C (retention time: 2.65; MS m/z 580 (M+H)⁺

Step 6:

To a solution of BocNH-P3 (t-BuGly)-P2[(Boc (4R)-(naphthal-1-oxo)proline)]-P1(1R,2S Vinyl Acca)-OH (Example 470, Step 5) (65 mgs, 0.11mmole) in 3.7 mL tetrahydrofuran was added 1,1′-carbonyl diimidazole (22mgs, 0.135 mmole).

The resulting mixture was refluxed for 30 minutes and then cooled toroom temperature. At this point, methanesulfonamide (27 mgs, 0.28 mmole)and DBU (34 L, 0.224 mmole) were added. The reaction was stirred for 2days and then more DBU (10 L) and methanesulfonamide (9 mgs) were added.After 24 hours, the reaction was diluted with 50 mL ethyl acetate andwashed with 50 mL 0.25N HCl and 50 mL brine. The solution was dried withsodium sulfate and concentrated in vacuo. The crude material waspurified by preparative TLC (3-2 ethyl acetate-hexanes) to give Compound470 (21 mgs, 28%) as a white filmy solid.

¹H NMR (d₄-MeOH, 500 MHz) δ 1.04 (s, 9H), 1.36 (s, 9H), 1.88 (t, 1H),2.18 (m, 1H), 2.31 (m, 1H), 2.63 (m, 1H), 3.11 (bs, 3H), 4.076 (m, 1H),4.30 (bd, 1H), 4.41 (bd, 1H), 4.52 (apparent t, 1H), 5.07 (m, 1H),5.24-5.30 (m, 2H), 5.80 (m, 1H), 6.92 (d, 1H, J=7.45 Hz), 7.35-7.46 (m,4H), 7.76 (d, 1H, J=8.1 Hz), 8.13 (d, 1H, J=8.3 Hz);

LC-MS C (retention time: 2.57; MS m/z 657 (M+H)⁺

Example 471 Preparation of Compound 471

Step 1:

To P2[Boc(4R)-(naphthal-1-oxo)proline]-P1(1S,2R Vinyl Acca)-OEt (Example470, Step 3, lower Rf) (91 mgs, 0.18 mmole) was added 4N HCl in dioxane(2.0 mL) and the solution was allowed to stir for 30 minutes.Concentration in vacuo yielded the HCl salt of P2[(4R)-(naphthal-1-oxo)proline)]-P1(1S,2R Vinyl Acca)-OEt as a yellow oil which was taken on tothe next step directly without further purification.

To a solution of N-Boc-L-tert-leucine-OH or BOC L-tBuGly (85 mgs, 0.37mmole) and the HCl salt of P2[(4R)-(naphthal-1-oxo) proline)]-P1(1S,2RVinyl Acca)-OEt (product obtained from reaction mentioned above) (0.18mmole) in 13 mL acetonitrile was added DIPEA (160 μL, 0.92 mmole) andHATU (154 mgs, 0.41 mmole). The resulting solution was stirred undernitrogen for 17 hours and then diluted with 100 mL ethyl acetate. Theorganic layer was washed with water, 1N NaHCO₃, 1N HCl, water and brine.The solution was then dried with sodium sulfate and concentrated invacuo to yield the title compound as a clear film (53 mgs, 47%).

¹H NMR (dl-CHCl₃, 500 MHz) δ 1.02 (s, 9H), 1.22 (t, 3H, J=7.0 Hz), 1.39(s, 9H), 1.47 (m, 1H), 1.88 (dd, 1H, J=8.0, 5.5 Hz), 2.07 (m, 1H), 2.42(m, 1H), 2.80 (dt, J=13.8, 6.0 Hz, 1H), 3.96 (m, 1H), 4.14 (m, 2H), 4.34(m, 2H), 4.77 (t, 1H, J=7.2 Hz), 5.09-5.33 (m, 3H), 5.72 (m, 1H), 6.82(d, 1H, J=7.6 Hz), 7.34-7.50 (m, 4H), 7.77 (d, 1H, J=8.0 Hz), 8.15 (d,1H, J=8.25 Hz);

LC-MS C (retention time: 2.81; MS m/z 608 (M+H)⁺

Step 2:

This product was prepared according procedure described in Example 470,Step 5 (5 mg, 10%), except using the product of Example 471, Step 1instead.

¹H NMR (d₄-MeOH, 500 MHz) δ 0.99 (s, 9H), 1.28 (m, 1H), 1.37 (s, 9H),1.60 (m, 1H), 2.06 (m, 1H), 2.28 (m, 1H), 2.66 (m, 1H), 3.91 (m, 1H),4.33 (m, 2H), 4.61 (bt, 1H), 4.97 (d, 1H, J=11.0 Hz), 5.19 (m 2H), 6.09(m, 1H), 6.88 (d, 1H, J=7.1 Hz), 7.35-7.46 (m, 4H), 7.78 (d, 1H, J=8.2Hz), 8.12 (d, 1H, J=8.3 Hz);

LC-MS C (retention time: 2.60; MS m/z 580 (M+H)⁺

Step 3:

To a solution of BocN-P3(L-tBuGly)-P2[(Boc (4R)-(naphthal-1-oxo)proline)]-P1(1S,2R Vinyl Acca)-COOH (38 mgs, 0.066 mmole) (Example 471,Step 2) in 2.2 mL tetrahydrofuran was added 1,1′-carbonyl diimidazole(13 mgs, 0.079 mmole). The resulting mixture was refluxed for 30 minutesand then cooled to room temperature. At this point, methanesulfonamide(16 mgs, 0.16 mmole) and DBU (20 μL, 0.13 mmole) were added. Thereaction was stirred for 2 days and then more DBU (10 μL) andmethanesulfonamide (9 mgs) were added. After 24 hours, the reaction wasdiluted with 50 mL ethyl acetate and washed with 50 mL 0.25N HCl and 50mL brine. The solution was dried with sodium sulfate and concentrated invacuo. The crude product was purified using one 20×40 cM preparative TLCplate from Analtech (eluent-3-2 ethyl acetate-hexanes) to give Compound471 (25 mgs, 58%) as a white filmy solid.

¹H NMR (d₄-MeOH, 500 MHz) δ 1.03 (s, 9H), 1.34 (s, 9H), 1.80 (m, 1H),2.18 (m, 1H), 2.31 (m, 1H), 2.68 (m, 1H), 3.09 (bs, 3H), 4.04 (m, 1H),4.20-4.44 (m, 2H), 4.51 (apparent t, 1H), 5.08 (m, 1H), 5.25-5.31 (m,2H), 5.77 (m, 1H), 6.93 (d, 1H, J=7.6 Hz), 7.36-7.45 (m, 4H), 7.77 (d,1H, J=8.0 Hz), 8.15 m, 1H);

LC-MS C (retention time: 2.57; MS m/z 657 (M+H)⁺

Section L Example 472 Biological Studies

Recombinant HCV NS3/4A Protease Complex FRET Peptide Assay

The purpose of this in vitro assay was to measure the inhibition of HCVNS3 protease complexes, derived from the BMS, H77C or J4l6S strains, asdescribed below, by compounds of the present invention. This assayprovides an indication of how effective compounds of the presentinvention would be in inhibiting HCV proteolytic activity.

Serum from an HCV-infected patient was obtained from Dr. T. Wright, SanFrancisco Hospital. An engineered full-length cDNA (complimentdeoxyribonucleic acid) template of the HCV genome (BMS strain) wasconstructed from DNA fragments obtained by reversetranscription-PCR(RT-PCR) of serum RNA (ribonucleic acid) and usingprimers selected on the basis of homology between other genotype 1astrains. From the determination of the entire genome sequence, agenotype 1a was assigned to the HCV isolate according to theclassification of Simmonds et al. (See P Simmonds, K A Rose, S Graham, SW Chan, F McOmish, B C Dow, E A Follett, P L Yap and H Marsden, J. Clin.Microbiol., 31(6), 1493-1503 (1993)). The amino acid sequence of thenonstructural region, NS2-5B, was shown to be >97% identical to HCVgenotype 1a (H77C) and 87% identical to genotype 1b (J4L6S). Theinfectious clones, H77C (1a genotype) and J4L6S (1b genotype) wereobtained from R. Purcell (NIH) and the sequences are published inGenbank (AAB67036, see Yanagi, M., Purcell, R. H., Emerson, S. U. andBukh, J. Proc. Natl. Acad. Sci. U.S.A. 94(16), 8738-8743 (1997);AF054247, see Yanagi, M., St Claire, M., Shapiro, M., Emerson, S. U.,Purcell, R. H. and Bukh, J, Virology 244(1), 161-172. (1998)).

The BMS, H77C and J4L6S strains were used for production of recombinantNS3/4A protease complexes. DNA encoding the recombinant HCV NS3/4Aprotease complex (amino acids 1027 to 1711) for these strains weremanipulated as described by P. Gallinari et al. (see Gallinari P,Paolini C, Brennan D, Nardi C, Steinkuhler C, De Francesco R.Biochemistry. 38(17):5620-32, (1999)). Briefly, a three-lysinesolubilizing tail was added at the 3′-end of the NS4A coding region. Thecysteine in the P1 position of the NS4A-NS4B cleavage site (amino acid1711) was changed to a glycine to avoid the proteolytic cleavage of thelysine tag. Furthermore, a cysteine to serine mutation was introduced byPCR at amino acid position 1454 to prevent the autolytic cleavage in theNS3 helicase domain. The variant DNA fragment was cloned in the pET21bbacterial expression vector (Novagen) and the NS3/4A complex wasexpressed in Escherichia coli strain BL21 (DE3) (Invitrogen) followingthe protocol described by P. Gallinari et al. (see Gallinari P, BrennanD, Nardi C, Brunetti M, Tomei L, Steinkuhler C, De Francesco R., J.Virol. 72(8):6758-69 (1998)) with modifications. Briefly, NS3/4Aexpression was induced with 0.5 mM Isopropyl β-D-1-thiogalactopyranoside(IPTG) for 22 hr at 20° C. A typical fermentation (10 L) yieldedapproximately 80 g of wet cell paste. The cells were resuspended inlysis buffer (10 mL/g) consisting of 25 mMN-(2-Hydroxyethyl)piperazine-N′-(2-Ethane Sulfonic acid) (HEPES), pH7.5,20% glycerol, 500 mM Sodium Chloride (NaCl), 0.5% Triton-×100, 1 ug/mllysozyme, 5 mM Magnesium Chloride (MgCl₂), 1 ug/ml Dnasel, 5 mMβ-Mercaptoethanol (βME), Protease inhibitor—Ethylenediamine Tetraaceticacid (EDTA) free (Roche), homogenized and incubated for 20 mins at 4° C.The homogenate was sonicated and clarified by ultra-centrifugation at235000 g for 1 hr at 4° C. Imidazole was added to the supernatant to afinal concentration of 15 mM and the pH adjusted to 8.0. The crudeprotein extract was loaded on a Nickel-Nitrilotriacetic acid (Ni-NTA)column pre-equilibrated with buffer B (25 mM HEPES, pH8.0, 20% glycerol,500 mM NaCl, 0.5% Triton-×100, 15 mM imidazole, 5 mM βME). The samplewas loaded at a flow rate of 1 mL/min. The column was washed with 15column volumes of buffer C (same as buffer B except with 0.2%Triton-×100). The protein was eluted with 5 column volumes of buffer D(same as buffer C except with 200 mM Imidazole).

NS3/4A protease complex-containing fractions were pooled and loaded on adesalting column Superdex-S200 pre-equilibrated with buffer D (25 mMHEPES, pH7.5, 20% glycerol, 300 mM NaCl, 0.2% Triton-×100, 10 mM βME).Sample was loaded at a flow rate of 1 mL/min. NS3/4A proteasecomplex-containing fractions were pooled and concentrated toapproximately 0.5 mg/ml. The purity of the NS3/4A protease complexes,derived from the BMS, H77C and J4L6S strains, were judged to be greaterthan 90% by SDS-PAGE and mass spectrometry analyses.

The enzyme was stored at −80° C., thawed on ice and diluted prior to usein assay buffer. The substrate used for the NS3/4A protease assay wasRET S1 (Resonance Energy Transfer Depsipeptide Substrate; AnaSpec, Inc.cat #22991)(FRET peptide), described by Taliani et al. in Anal. Biochem.240(2):60-67 (1996). The sequence of this peptide is loosely based onthe NS4A/NS4B natural cleavage site except there is an ester linkagerather than an amide bond at the cleavage site. The peptide substratewas incubated with one of the three recombinant NS3/4A complexes, in theabsence or presence of a compound of the present invention, and theformation of fluorescent reaction product was followed in real timeusing a Cytofluor Series 4000.

The reagents were as follow: HEPES and Glycerol (Ultrapure) wereobtained from GIBCO-BRL. Dimethyl Sulfoxide (DMSO) was obtained fromSigma. β-Mercaptoethanol was obtained from Bio Rad.

Assay buffer: 50 mM HEPES, pH7.5; 0.15M NaCl; 0.1% Triton; 15% Glycerol;10 mM βME. Substrate: 2 μM final concentration (from a 2 mM stocksolution in DMSO stored at −20° C.). HCV NS3/4A type 1a (1b), 2-3 nMfinal concentration (from a 5 μM stock solution in 25 mM HEPES, pH7.5,20% glycerol, 300 mM NaCl, 0.2% Triton-×100, 10 mM βME).

The assay was performed in a 96-well polystyrene black plate fromFalcon. Each well contained 25 μl NS3/4A protease complex in assaybuffer, 50 μl of a compound of the present invention in 10% DMSO/assaybuffer and 25 μl substrate in assay buffer. A control (no compound) wasalso prepared on the same assay plate. The enzyme complex was mixed withcompound or control solution for 1 min before initiating the enzymaticreaction by the addition of substrate. The assay plate was readimmediately using the Cytofluor Series 4000 (Perspective Biosystems).The instrument was set to read an emission of 340 nm and excitation of490 nm at 25° C. Reactions were generally followed for approximately 15minutes.

The percent inhibition was calculated with the following equation:100−[(δF _(inh) /δF _(con))×100]where δF is the change in fluorescence over the linear range of thecurve. A non-linear curve fit was applied to theinhibition-concentration data, and the 50% effective concentration(IC₅₀) was calculated by the use of Excel XI-fit software using theequation, y=A+((B−A)/(1+((C/x)^D))).

All of the compounds tested were found to have IC50s of 10 μM or less.Further, compounds of the present invention, which were tested againstmore than one type of NS3/4A complex, were found to have similarinhibitory properties though the compounds uniformly demonstratedgreater potency against the 1b strains as compared to the 1a strains.

Specificity Assays

The specificity assays were performed to demonstrate the selectivity ofthe compounds of the present invention in inhibiting HCV NS3/4A proteaseas compared to other serine or cysteine proteases.

The specificities of compounds of the present invention were determinedagainst a variety of serine proteases: human leukocyte elastase (HLE),porcine pancreatic elastase (PPE) and human pancreatic chymotrypsin andone cysteine protease: human liver cathepsin B. In all cases a 96-wellplate format protocol using colorimetric p-nitroaniline (pNA) substratespecific for each enzyme was used as described previously (Patent WO00/09543) with some modifications to the serine protease assays. Allenzymes were purchased from Sigma while the substrates were from Bachem.

Each assay included a 2 hr enzyme-inhibitor pre-incubation at RTfollowed by addition of substrate and hydrolysis to ˜30% conversion asmeasured on a Spectramax Pro microplate reader. Compound concentrationsvaried from 100 to 0.4 μM depending on their potency.

The final conditions for each assay were as follows:

50 mM Tris(hydroxymethyl)aminomethane hydrochloride (Tris-HCl) pH8, 0.5MSodium Sulfate (Na₂SO₄), 50 mM NaCl, 0.1 mM EDTA, 3% DMSO, 0.01%Tween-20 with:

133 μM succ-AAA-pNA and 20 nM HNE or 8 nM PPE; 133 μM succ-AAV-pNA and15 nM HLE; 100 μM succ-AAPF-pNA and 250 μM Chymotrypsin.

100 mM NaHPO₄ (Sodium Hydrogen Phosphate) pH 6, 0.1 mM EDTA, 3% DMSO, 1mM TCEP (Tris(2-carboxyethyl)phosphine hydrochloride), 0.01% Tween-20,30 μM Z-FR-pNA and 5 nM Cathepsin B (enzyme stock activated in buffercontaining 20 mM TCEP before use).

The percentage of inhibition was calculated using the formula:[1−((UV_(inh)−UV_(blank))/(UV_(ctl)−UV_(blank)))]×100

A non-linear curve fit was applied to the inhibition-concentration data,and the 50% effective concentration (IC₅₀) was calculated by the use ofExcel Xl-fit software.

HCV Replicon Cell-Based Assay

An HCV replicon whole cell system was established as described byLohmann V, Korner F, Koch J, Herian U, Theilmann L, Bartenschlager R.,Science 285(5424): 110-3 (1999). This system enabled us to evaluate theeffects of our HCV Protease compounds on HCV RNA replication. Briefly,using the HCV strain 1B sequence described in the Lohmann paper(Accession number: AJ238799), an HCV cDNA was generated encoding the 5′internal ribosome entry site (IRES), the neomycin resistance gene, theEMCV (encephalomyocarditis virus)-IRES and the HCV nonstructuralproteins, NS3-NS5B, and 3′ non-translated region (NTR). In vitrotranscripts of the cDNA were transfected into the human hepatoma cellline, Huh7. Selection for cells constitutively expressing the HCVreplicon was achieved in the presence of the selectable marker, neomycin(G418). Resulting cell lines were characterized for positive andnegative strand RNA production and protein production over time.

Huh7 cells, constitutively expressing the HCV replicon, were grown inDulbecco's Modified Eagle Media (DMEM) containing 10% Fetal calf serum(FCS) and 1 mg/ml G418 (Gibco-BRL). Cells were seeded the night before(1.5×10⁴ cells/well) in 96-well tissue-culture sterile plates. Compoundand no compound controls were prepared in DMEM containing 4% FCS, 1:100Penicillin/Streptomysin, 1:100 L-glutamine and 5% DMSO in the dilutionplate (0.5% DMSO final concentration in the assay). Compound/DMSO mixeswere added to the cells and incubated for 4 days at 37° C. After 4 days,plates were rinsed thoroughly with Phosphate-Buffered Saline (PBS) (3times 150 μl). The cells were lysed with 25 μl of a lysis assay reagentcontaining the FRET peptide (RET S1, as described for the in vitroenzyme assay). The lysis assay reagent was made from 5× cell Luciferasecell culture lysis reagent (Promega #E153A) diluted to 1× with distilledwater, NaCl added to 150 mM final, the FRET peptide diluted to 10 μMfinal from a 2 mM stock in 100% DMSO. The plate was then placed into theCytofluor 4000 instrument which had been set to 340 nm excitation/490emission, automatic mode for 21 cycles and the plate read in a kineticmode. EC₅₀ determinations were carried out as described for the IC₅₀determinations.

As a secondary assay, EC₅₀ determinations from the replicon FRET assaywere confirmed in a quantitative RNA assay. Cells were lyzed using theRneasy kit (Qiagen). Purified total RNA was normalized using RiboGreen(Jones L J, Yue S T, Cheung C Y, Singer V L, Anal. Chem., 265(2):368-74(1998)) and relative quantitation of HCV RNA expression assessed usingthe Taqman procedure (Kolykhalov A A, Mihalik K, Feinstone S M, Rice CM, Journal of Virology 74, 2046-2051 (2000)) and the PlatinumQuantitative RT-PCR Thermoscript One-Step kit (Invitrogen cat#11731-015). Briefly, RNA made to a volume of 5 μl (≦1 ng) was added toa 20 μl Ready-Mix containing the following: 1.25× Thermoscript reactionmix (containing Magnesium Sulfate and 2-deoxynucleoside 5′-triphosphates(dNTPs)), 3 mM dNTPs, 200 nM forward primer (sequence:5′-gggagagccatagtggtctgc-3′), 600 nM reverse primer(5′-cccaaatctccaggcattga-3′), 100 nM probe(5′-6-FAM-cggaattgccaggacgaccgg-BHQ-1-3′)(FAM: Fluorescein-aminohexylamidite; BHQ: Black Hole Quencher), 1 μM Rox reference dye (Invitrogencat #12223-012) and Thermoscript Plus Platinum Taq polymerase mixture.All primers were designed with ABI Prism 7700 software and obtained fromBiosearch Technologies, Novato, Calif. Samples containing knownconcentrations of HCV RNA transcript were run as standards. Using thefollowing cycling protocol (50° C., 30 min; 95° C., 5 min; 40 cycles of95° C., 15 sec, 60° C., 1 min), HCV RNA expression was quantitated asdescribed in the Perkin Elmer manual using the ABI Prism 7700 SequenceDetector.

Biological Examples

Representative compounds of the invention were assessed in the HCVreplicon cell assay and/or in several of the outlined specificityassays. For example, Compound 34 was found to have an IC₅₀ of 23nanomolar (nM) against the NS3/4A BMS strain in the enzyme assay.Similar potency values were obtained with the published H77C (IC₅₀ of 3nM) and J4L6S (IC₅₀ of 2.9 nM) strains. The EC₅₀ value in the repliconassay was 166 nM.

In the specificity assays, the same compound was found to have thefollowing activity: HLE>100 μM; PPE>200 μM; Chymotrypsin>200 μM;Cathepsin B>200 μM. These results indicate this family of compounds arehighly specific for the NS3 protease and many of these members inhibitHCV replicon replication.

The compounds of the current invention were tested and found to haveactivities in the ranges as follow:

IC50 Activity Ranges (NS3/4A BMS Strain): A is 10-100 micromolar (μM); Bis 1-10 μM; C is 0.1-1 μM; D is <0.1 μM

EC50 Activity Range (for compounds tested): A is 10-100 μM; B is 1-10μM; C is 0.1-1 μM; D is <0.1 μM

Note that by using the Patent example number and the Patent compoundnumber shown in the table the structures of compounds can be foundherein.

In accordance with the present invention, preferably the compounds havea biological activity (EC₅₀) of 10 μM or less, more preferably 1 μM orless and most preferably 100 nM or less.

TABLE 1 Biological Activity Patent Patent IC50 EC50 Example Cmpd rangerange Number Number D D 1 1 D C 2 2 D D 3 3 D D 4 4 D D 5 5 C C 6 6 D C7 7 C B 8 8 D C 9 9 D D 10 10 D D 11 11 D D 12 12 D D 13 13 D D 14 14 CB 15 15 D C 16 16 D D 17 17 D D 18 18 D D 19 19 D C 20 20 D D 21 21 D C22 22 D D 23 23 D D 24 24 D D 25 25 D D 26 26 D D 27 27 D D 28 28 D D 2929 D D 30 30 D D 31 31 D D 32 32 D D 33 33 D C 34 34 D D 35 35 D D 36 36D D 37 37 D D 38 38 D D 39 39 D C 40 40 D D 41 41 D D 42 42 C B 45 45 CB 46 46 D C 47 47 B 48 48 B 49 49 C B 50 50 B 52 52 C B 53 53 D C 55 55D D 56 56 D C 57 57 D D 58 58 D C 59 59 D B 60 60 C B 61 61 D D 62 62 DB 63 63 D D 64 64 D C 65 65 D C 66 66 D D 67 67 D D 68 68 D D 69 69 D D70 70 D D 71 71 D C 72 72 D C 73 73 D D 74 74 D D 75 75 D D 76 76 D C 7777 D B 78 78 D C 79 79 D B 80 80 D C 81 81 D C 82 82 D C 83 83 D C 84 84C B 85 85 B A 86 86 B A 87 87 B A 88 88 D D 89 89 D C 91 91 D D 92 92 CC 93 93 D D 94 94 D C 95 95 D D 96 96 D D 97 97 B 99 99 C B 100 100 D D101 101 D D 102 102 D D 103 103 D D 104 104 D D 105 105 D D 106 106 D C107 107 D D 108 108 D D 109 109 D C 110 110 D C 120 120 D C 121 121 D C122 122 D C 123 123 C B 124 124 D D 125 125 D C 126 126 D C 127 127 D C128 128 C C 129 129 D C 130 130 C B 131 131 D B 132 132 C B 133 133 D C134 134 D C 135 135 D C 136 136 D D 137 137 D D 138 138 D C 139 139 D B140 140 D D 141 141 D D 142 142 D D 143 143 D C 144 144 D C 145 145 D D146 146 D B 147 147 D D 148 148 D D 149 149 D D 150 150 D D 151 151 D D152 152 D D 153 153 D C 154 154 D D 155 155 D C 180 180 D C 181 181 D C182 182 D D 183 183 D D 185 185 D D 186 186 D C 187 187 D C 188 188 D C189 189 D C 190 190 D C 191 191 D D 192 192 D C 193 193 D C 194 194 D D195 195 D B 196 196 C A 197 197 D C 198 198 D C 199 199 D D 200 200 D D201 201 D B 202 202 D C 204 204 D D 206 206 D D 207 207 D C 209 209 D D210 210 D D 211 211 D D 212 212 D D 213 213 D D 215 215 D D 219 219 D D220 220 D D 223 223 D C 224 224 D C 225 225 D D 227 227 D D 229 229 C C230 230 B 231 231 D D 232 232 C C 233 233 D C 235 235 D C 237 237 D C238 238 D D 239 239 D D 240 240 D D 241 241 D B 242 242 C A 243 243 D A244 244 C 245 245 D D 250 250 D D 251 251 D D 252 252 D D 253 253 D D254 254 D C 255 255 A 256 256 D D 257 257 D D 258 258 D D 259 259 D C260 260 D D 261 261 D D 262 262 C C 263 263 C B 264 264 D D 265 265 D D266 266 D D 267 267 D D 268 268 D C 269 269 D C 270 270 D C 271 271 D D272 272 D D 273 273 D C 274 274 D D 275 275 D D 276 276 D D 277 277 D D278 278 D D 279 279 D D 280 280 D D 281 281 D C 282 282 D C 283 283 C A284 284 D D 285 285 D D 286 286 D C 287 287 D B 288 288 D C 289 289 D D290 290 D D 291 291 D C 292 292 D C 293 293 D C 294 294 C B 295 295 D C296 296 C B 297 297 D D 298 298 D C 299 299 D C 300 300 D D 320 320 D C321 321 D D 322 322 D C 323 323 D C 324 324 D D 325 325 D D 326 326 D D327 327 D D 328 328 D D 329 329 D D 330 330 D 331 331 C C 334 334 D B335 335 C B 336 336 C B 337 337 D B 338 338 D B 339 339 D A 340 340 B341 341 B 342 342 B 343 343 C 344 344 B A 345 345 B 346 346 C A 347 347A 348 348 B 349 349 C A 350 350 B 351 351 A 352 352 A 353 353 B 354 354C A 355 355 C B 356 356 D C 357 357 C B 370 370 D D 371 371 D D 372 372D D 373 373 D C 374 374 D D 375 375 D D 376 376 D C 377 377 D D 378 378D D 379 379 D C 380 380 D C 381 381 D C 382 382 D D 383 383 D D 384 384D D 385 385 C B 386 386 D D 410 410 D C 411 411 D D 412 412 D D 413 413D D 414 414 D D 415 415 D C 420 420 D C 421 421 D B 422 422 C 423 423 DC 424 424 C B 425 425 D C 426 426 D C 427 427 D C 428 428 D D 429 429 DC 430 430 D C 431 431 D C 432 432 C B 433 433 D C 434 434 D C 435 435 DC 436 436 D C 437 437 D C 438 438 D C 450 450 B 451 451 B 452 452 453453 C B 454 454 C 470 470 B 471 471

Section M

The following compounds that can be made using the methods describedherein and specifically in sections A through K of the exemplificationsection and more specifically in sections B, E, F and G. Moreover itshould be made clear that each of the groups B, R₃, R₂ and R₁ shownbelow can be replaced by any of the groups exemplified in sections Athrough K and elsewhere herein or designated in Formula I. For examplethe R₃ group in Table 2 is shown as a t-butyl group but one skilled inthe art would recognize that for each of the entries cited below thisgroup could be replaced with an isopropyl group or a C₁₋₆ alkylsubstituted with an alkoxy. Or the B group shown below could be replacedwith a tert-butyl urea moiety for each of the entries cited below.

TABLE 2

C(1,2)- Cyclo- propane En- stereo- try B R₃ X R′ R₂ chemistry R₁ A

O

(1R, 2S)

B

O

(1R, 2S)

C

O

(1R, 2S)

D

O

(1R, 2S)

E

O

(1R, 2S)

F

O

(1R, 2S)

G

O

(1R, 2S)

H

O

(1R, 2S)

I

O

(1R, 2S)

J

O

(1R, 2S)

K

O

(1R, 2S)

L

O

(1R, 2S)

M

O

(1R, 2S)

N

O

(1R, 2S)

O

O

(1R, 2S)

P

O

(1R, 2S)

Q

O

(1R, 2S)

R

O

(1R, 2S)

S

O

(1R, 2S)

T

O

(1R, 2S)

U

O

(1R, 2S)

V

O

(1R, 2S)

W

O

(1R, 2S)

X

O

(1R, 2S)

Y

O

(1R, 2S)

Z

O

(1R, 2S)

Aa

O

(1R, 2S)

Bb

O

(1R, 2S)

Cc

O

(1R, 2S)

Dd

O

(1R, 2S)

Ee

O

(1R, 2S)

Ff

O

(1R, 2S)

Gg

O

(1R, 2S)

Hh

O

(1R, 2S)

Ii

O

(1R, 2S)

Jj

O

(1R, 2S)

Kk

O

(1R, 2S)

Ll

O

(1R, 2S)

Mm

O

(1R, 2S)

What is claimed is:
 1. A compound which is:

or a salt thereof.